+

WO2016005971A1 - Dispositifs pour la culture d'un échantillon biologique dans un liquide de culture - Google Patents

Dispositifs pour la culture d'un échantillon biologique dans un liquide de culture Download PDF

Info

Publication number
WO2016005971A1
WO2016005971A1 PCT/IL2015/050698 IL2015050698W WO2016005971A1 WO 2016005971 A1 WO2016005971 A1 WO 2016005971A1 IL 2015050698 W IL2015050698 W IL 2015050698W WO 2016005971 A1 WO2016005971 A1 WO 2016005971A1
Authority
WO
WIPO (PCT)
Prior art keywords
culturing
fluid
biological sample
module
chamber
Prior art date
Application number
PCT/IL2015/050698
Other languages
English (en)
Inventor
Amir Arav
Original Assignee
Fertilesafe Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fertilesafe Ltd filed Critical Fertilesafe Ltd
Publication of WO2016005971A1 publication Critical patent/WO2016005971A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/10—Perfusion
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C12M41/16—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature by recirculation of culture medium at controlled temperature

Definitions

  • the invention relates to the field of tissue cultures. More precisely, the invention relates to devices for culturing tissues.
  • Culturing biological samples is long known in the art.
  • tissue culturing one example is in-vitro fertilization (known as IVF), wherein the embryo is grown, cultured, for several days (e.g Thompson four days) until it reaches the a blastocyst stage, whereupon it is implanted in a uterus.
  • IVF in-vitro fertilization
  • the present invention in some embodiments thereof, relates to culturing a biological sample.
  • the invention relates to a device for culturing a biological sample in a culturing fluid, the device comprising:
  • a culturing module configured to hold a first portion of the culturing fluid and to allow culturing the biological sample in the first portion;
  • a chamber coupleable to the culturing module via a fluidic connection, the chamber is configured to hold a second portion of the culturing fluid in a hollow space of the chamber, to receive flowing culturing fluid which flows from the culturing module via the fluidic connection, and to hold the flowing culturing fluid within the hollow space as part of the second portion ;
  • a circulator coupled to the hollow space of the chamber, configured to drain a circulated culturing fluid from the culturing fluid in the second portion and to convey the circulated culturing fluid to the first portion in the culturing module; wherein the device is configured to isolate the culturing fluid and the biological sample from direct contact with an environment external to the device.
  • the culturing module is configured to hold a permeable member, thereby preventing the biological sample from flowing out of the culturing module.
  • the permeable member is a removable permeable member.
  • the permeable member is an openable permeable member.
  • the culturing module is configured to hold a carrier for culturing the biological sample, wherein the carrier is configured to hold the biological sample submersed within the first portion of the culturing fluid.
  • the invention relates to a device for culturing a biological sample in a culturing fluid, the device comprising:
  • a first container configured to hold a first portion of the culturing fluid and to allow culturing the biological sample in the first portion, wherein the first portion comprises culturing fluid received as in-flow;
  • a second container configured to hold a second portion of the culturing fluid wherein the second portion comprises culturing fluid received as out-flow from the first portion.
  • the device is configured to isolate the culturing fluid and the biological sample from direct contact with an environment external to the device.
  • the first container is enclosed within the second container.
  • the first container is external to the second container.
  • the first container is coupleable to the second container via a closed path allowing flow of fluids.
  • the invention relates to a device for culturing a biological sample in a culturing fluid, the device comprising:
  • a culturing module configured to hold a first portion of the culturing fluid and to allow culturing the biological sample in the first portion, wherein the culturing fluid in the first portion is substantially in a physiological temperature of the biological sample; a chamber configured to hold a second portion of the culturing fluid in a hollow space of the chamber wherein the culturing fluid in the second portion is colder than the temperature of the fluid in the first portion.
  • FIG 1 schematically illustrates a device for culturing a biological sample, according to some embodiments of the invention
  • FIG 2A schematically illustrates a device for culturing a biological sample, according to some embodiments of the invention
  • FIG 2B schematically illustrates a device for culturing a biological sample, according to some embodiments of the invention
  • FIG 2C schematically illustrates a culturing module for culturing a biological sample, according to some embodiments of the invention
  • FIG 2D schematically illustrates a culturing module for culturing a biological sample, according to some embodiments of the invention
  • FIG 3 illustrates an upper view of a device for culturing a plurality of biological samples, according to some embodiments of the invention
  • FIG 4 illustrates a side view of a device for culturing a plurality of biological samples, according to some embodiments of the invention
  • FIG 5 schematically illustrates embodiments of the invention wherein a pump is used to drive flow, according to some embodiments of the invention
  • FIG 6 schematically illustrates a device comprising a cooled chamber and a culturing module coupled via a fluidic path, according to some embodiments of the invention
  • FIG 7 schematically illustrates a device for culturing a biological sample configured to submerge the sample in a warm culturing fluid
  • FIG 8 provides a device for culturing a biological sample configured to submerge the sample in a warm culturing fluid, according to some embodiments of the invention.
  • FIG 9 schematically illustrates a device for culturing a biological sample, according to some embodiments of the invention.
  • the description herein discloses embodiments of devices for culturing biological samples.
  • the term “device” is analog to "a device for culturing a biological sample” or to "a culturing device”.
  • sample refers to "a biological sample”, including “a reproductive biological sample”, and specifically including “an oocyte”, “sperm”, “an embryo” etc.
  • biological sample may include also any other, non- reproductive biological sample subject to culturing in accordance with embodiments of the invention.
  • FIG 1 schematically illustrates a device 100 for culturing a biological sample
  • the device may be utilized, for example, for in-vitro fertilization of an oocyte by a sperm, and/or for in-vitro culturing an embryo prior to implantation (e.g., until a blastocyst is achieved), or for culturing of any other a biological sample as mentioned above.
  • device 100 is configured to hold a biological sample 101 submerged within a culturing fluid 106.
  • the culturing fluid is configured to provide appropriate conditions for culturing the biological sample, such as composition, pH (measure of the acidity of the fluid), etc.
  • FIG 1 illustrates embodiments of device 100 that comprise a chamber 103, within which there is a culturing module 104 that holds a carrier 102. Therefore, device 100 comprises a chamber 103 and a culturing module 104.
  • the culturing module is configured to hold a "first portion of the culturing fluid" 108, shortly referred to as a "first portion”.
  • culturing module 104 is configured to hold a carrier 102, which in turn is configured to culture the sample.
  • Chamber 103 has an opening 105, through which carrier 102 protrudes, while the carrier closes and seals the opening in order to allow keeping the culturing fluid and the sample sterile.
  • opening 105 may be used for placing carrier 102 in culturing module 104.
  • Carrier 102 may have a permeable member 107, e.g., at the bottom thereof.
  • the permeable member is configured to enable a thru-flow of the culturing fluid between sides thereof, while preventing the biological sample from being carried along with the thru-flow.
  • carrier 102 is configured to hold the biological sample submersed within the culturing fluid that is held in the culturing module.
  • the culturing module is configured to allow culturing the biological sample in the first portion of the culturing fluid.
  • FIG 1 are non-limiting and in other embodiments the culturing module and the carrier may be internal to the chamber 2A01, as illustrated by device 2A00 of FIG 2A. It should be appreciated that the carrier 102 in such embodiments should be placed inside chamber 2A01 for culturing biological sample 101. Placing the carrier may be enabled, for example, by a chamber having an openable lid 2A02, as illustrated in FIG 2A, which seals the chamber when closed, or by any other mean alternative or additional to lid 2A02. It is noted that apart from the opening mechanism, chamber 2A01 resembles chamber 103 of FIG 1 and therefore hereinafter reference to chamber 103 refers to chamber 2A01 as well, unless specifically noted.
  • device 2B00 comprises a culturing module 2B01, which is external to chamber 2B02.
  • culturing device 2B01 is also configured to store a first portion of the culturing fluid 108 and to facilitate culturing the biological sample 101 in the first portion of the culturing fluid. More specifically, culturing module 2B01 is configured to facilitate culturing a biological sample 101 by holding carrier 102.
  • embodiments of the invention comprise a culturing module (such as 104 and 2B01), which is configured to store a first portion 108 of culturing fluid 106.
  • the culturing module is configured also to facilitate culturing biological sample 101 in the first portion of the culturing fluid.
  • the culturing modules are configured to hold carrier 102 for culturing the biological sample, wherein the carrier is configured to hold the sample submersed within the first portion of the culturing fluid.
  • culturing may be done without a carrier.
  • FIG 2C schematically illustrates a culturing module 2C00 for culturing a biological sample 2C01, according to some embodiments of the invention.
  • culturing modules such as 2C00 culturing may be done directly in the culturing module.
  • Such embodiments are applicable, e.g. when the biological sample is large enough (for example, above 0.25 millimeter in radius), hence it cannot be lost in the volume of first portion 108.
  • culturing fluid flows out of the culturing module, and therefore, in embodiments such as those depicted in FIG 2C it is required to prevent the sample from flowing out of culturing module 2C00.
  • outflow must occur via an opening in culturing module 2C00, such prevention can be made, for example, by placing a permeable obstacle 2C02 between biological sample 2C01 and the opening, or at the opening.
  • outflow occurs when the culturing fluid overflows from culturing module 104.
  • FIG 2C Such an example is illustrated also in FIG 2C, wherein overflow occurs above a rim 2C03 of culturing module 2C00. Therefore, the opening through which outflow occurs is the rim of culturing module 2C00 or possibly the lowest place along the rim. The height of the lowest place is schematically marked by 2C04 in the figure.
  • a permeable obstacle 2C02 e.g., in a way closing the opening within the rim, at a level 2C04 of the lowest place along the rim, or possibly below that level.
  • culturing module 2C00 may be configured to have a removable permeable obstacle. Additionally or alternatively, culturing module 2C00 may be coupleable to an openable permeable obstacle, e.g., by a pivot, a hinge, etc. In such embodiments, wherein the permeable obstacle is removable and/or openable, it is possible to place the permeable obstacle (and/or to close an opening therein) after placing the biological sample in the culturing module.
  • culturing modules such as 2C00 may be configured to hold a permeable obstacle, thereby preventing the biological sample from flowing out of the culturing module.
  • FIG 2D schematically illustrates a culturing module 2D00 for culturing a biological sample 2C01, according to some embodiments of the invention. Similar to the embodiments illustrated in FIG 2C, the sample in these embodiments is cultured directly in the culturing module. As explained before with reference to FIG. 2C, such embodiments are applicable for is large enough samples that cannot be lost in the volume of the first portion 108.
  • a permeable obstacle 2D01 is placed at the outlet, preventing the sample 2C01 from flowing out of culturing module 2D00. That is, similar to culturing module 2C00, culturing modules such as 2D00 may be configured to hold a permeable obstacle, thereby preventing the biological sample from flowing out of the culturing module.
  • a conduit is a pipe or channel for conveying fluids.
  • a conduit is therefore a hollow object used to conduct fluids.
  • Conduits may have herein any applicable form of cross section, such as cylindrical, square, triangular etc.
  • the cross section may even be irregular, i.e., non-symmetrical or departing from any defined/known form.
  • a conduit is not limited herein to a hollow object having the same cross section along its length, i.e., if applicable, the form of the cross section may change along the conduit. For example, in one point the cross section may be cylindrical, in another point it may rectangular, while in yet another point is may be irregular.
  • carrier 102 is configured to hold a sample submersed within the first portion of the culturing fluid so it can be cultured.
  • secretions it should be considered that by producing, giving rise to a flow of the culturing fluid around the sample, it is possible to move the secretions away from the sample, thereby improving the likelihood of the sample to survive.
  • Flow should be produced at least around the sample, that is, at least in the smaller amongst a volume of culturing fluid where secretions may accumulate around the sample and a volume where secretions may damage the sample. Therefore, according to embodiments of the invention, it is desired to produce flow of the culturing fluid around the sample.
  • the culturing fluid around the sample form part of the culturing fluid held by the carrier. Accordingly, by producing, flow of the culturing fluid held by the carrier, flow is produced around the sample as well.
  • a conveyed culturing fluid constitutes a "received fluid".
  • received fluid is received by the culturing fluid held by carrier 102 from conduit 106, coupled to carrier 102 via inlet 109.
  • the received fluid, marked 110 is schematically represented in the figure by the small winding arrows below inlet 109. Because carrier 102 comprises a permeable member at its bottom, at least part of received liquid 110 may flow therethrough.
  • the received liquid forms part of first portion 108 in culturing module 102, that is, the received fluid will enlarge the volume of first portion 108.
  • culturing module 102 has a rim, similar to culturing module 2C00 in FIG 2C, described above. Due to enlarging the volume of the first portion 108, at some point it may reach the rim, or a lowest part thereof, whereupon culturing fluid will start flowing out of culturing module 104 as overflow. Culturing fluid flowing out of culturing module 104 constitutes "fluid flow-out". In FIG 1 the fluid flow-out of culturing module 104 is marked 111, and is schematically represented by the small winding arrows down right of culturing module 104.
  • the carriers in these embodiments are held by the culturing modules, and that the carriers are configured to submerge the samples in the first portions. Accordingly, it is can be understood that by conveying culturing fluid to a first portion held by a culturing module, it is possible to produce flow of culturing fluid in the first portion, and therefore to produce flow around a sample submerged in the first portion.
  • FIG 2C the inlet is unseen though it can be appreciated that the culturing module 2C00 can be comprised in the devices of FIGs 1 and 2A. Therefore it is clear now that by conveying culturing fluid to first portion 108 held by culturing module 2C00, it is possible to produce flow of culturing fluid in the first portion, and therefore to produce flow around sample 2C01 submerged therein. This explanation applies also to culturing device 2D00 depicted in FIG 2D, mutatis mutandis.
  • the figure illustrates the existence of a hollow space 112 within the chamber.
  • the hollow space should be at a least partially lower than the opening of culturing module 104, from where the fluid flow- out is flowing.
  • chamber 103 holds culturing fluid in its bottom, and more specifically, in the bottom of its hollow space 112.
  • the culturing fluid held in hollow space 112 of chamber 103 constitutes a "second portion" 113 of the culturing fluid.
  • chamber 103 receives the fluid flow-out 111 coming from culturing module 104.
  • the fluid flow-out 111 of culturing module 104 serves as received fluid for chamber 103.
  • this flow 111 is enabled by a height difference existing between the opening in culturing module 104 from which the fluid flow-out is flowing, and between the level of second portion 113.
  • This height difference is represented in FIG 1 by the reference numeral 114. Accordingly, it may be appreciated that height difference 114 serves as coupling between the chamber and the culturing module.
  • culturing module 104 and second portion 113 are coupled, and when this flow is inactive, they may be considered as uncoupled (though they are coupleable as the flow may be re-activated). Height difference 114 therefore constitutes a "fluidic connection", while culturing module 104 and second portion 113 are coupleable via fluidic connection 114. Further to understanding this it can be appreciated the chamber is configured to receive flowing culturing fluid 111 via fluidic connection 114.
  • the culturing fluid received from the flow is held in hollow space 112, wherein it forms part of second portion 113. That is, chamber 103 is configured to hold the flowing culturing fluid within the hollow space as part of second portion 113 of the culturing fluid.
  • FIG 2B02 that also comprises a chamber 2B01 and a culturing module 2B01.
  • the culturing module 2B01 does not occupy space within chamber 2B02.
  • Chamber 2B02 confines and comprises a hollow space 2B03, however, culturing module 2B01 is external thereto.
  • the two elements namely culturing module 2B01 and chamber 2B02, may be coupled via a conduit 2B04, which forms a fluidic connection: flow of culturing fluid therethrough may be activated or deactivated, e.g., in accordance with a level of fluid in first portion 108 held by culturing module 2B01. Therefore, and further to understanding the embodiments of the invention presented with reference to FIG 1 and 2A, it should be understood that similarly to chambers 103 and 2A01, chamber 2B02 is coupleable as well to culturing module 2B01 via fluidic connection 2B04, while the chamber is configured to hold a second portion 113 of the culturing fluid in a hollow space 2B03 of the chamber.
  • the chamber is also configured to receive flowing culturing fluid which flows from culturing module 2B01 via the fluidic connection 2B04, and it is further configured to hold the flowing culturing fluid within hollow space 2B03 as part of second portion 113.
  • Circulator 115 appear in all three figures.
  • the circulator is a conduit coupled to hollow space 112 (in FIGs 1 and 2A) or 2B03 (in FIG 2B) of chamber 103 (in FIG 1), 2A01 (in FIG 2A) or 2B02 (in FIG 2B), and to the culturing module 104 (in FIGs 1 and 2A) or 2B01 (in FIG 2B).
  • the circulator is configured to drain a circulated culturing fluid 116 from the culturing fluid in the second portion 113, and to convey the circulated culturing fluid 116 to the first portion 108 in the culturing module 104 (in FIGs 1 and 2A) or 2B01 (in FIG 2B).
  • the circulated fluid residing at a certain point of time in the hollow space of circulator 115 constitute a "circulated portion of culturing fluid", or shortly a "circulated portion”.
  • the reader should understand the culturing fluid flowing at a certain point to time in the received fluid 110 of culturing module 102 has a certain volume, hence this fluid constitutes a "first in-flow portion of culturing fluid" or shortly a "first inflow portion".
  • the volume of the first in-flow portion may be small and negligible in certain cases, though the reader should be conscious of its existence.
  • the fluid flow-out 111 of culturing module 104 has a volume as well and therefore it constitutes a "first out-flow portion of culturing fluid" or shortly a "first out-flow portion".
  • the volume of the first out-flow portion may be small and negligible in certain cases, though the reader should be conscious of its existence as well.
  • the culturing modules describes with reference to FIGs 1-2D may be device into two types: "closed culturing module", such as the culturing modules 2B01 and 2D00, schematically illustrated in FIGs 2B and 2B, and "open culturing modules", such as culturing modules 104 and 2C00, schematically illustrated in FIGs 1, 2A and 2C.
  • closed culturing modules are always enclosed within chambers
  • closed culturing modules are always external to the chambers.
  • an open culturing module may be used with or without additional elements, and enclosed within a dedicated container.
  • the culturing module and the chamber may be external one to the other, is depicted in FIG 2B, however, instead of being implemented as two separated elements 2B01 and 2B02, coupled via a conduit 2B04, the two elements may be implemented in a single structure that forms the device.
  • FIG 4 example for such embodiments is depicted in FIG 4, described below.
  • FIG 3 illustrates an upper view of a device 300 for culturing a plurality of biological samples, according to some embodiments of the invention.
  • the depicted device is configured to culture three (3) samples, though this is non-limiting and other number of samples may be used if and when applicable. Therefore, it is generally noted that the device is configured to culture a plurality of samples.
  • Device 300 comprises three culturing modules 301, while each culturing module 301 is configured to hold a carrier 302. While in the figure all three carriers are depicted, it should be realized that device 300 can be operated when only part of the culturing modules hold a carrier, while others are vacant.
  • the device comprises a single fluid entrance 303, coupleable to a conduit 304, such as circulator 2B04 depicted in FIGs 1, 2A and 2B. Only the end of conduit 304 is depicted in the figure.
  • fluids enter and flow in a fluid path 305, that in the illustrated embodiment appear to be an annular fluid path or tunnel.
  • Splits 306 in path 305 lead portions of the flowing fluids towards carriers 302.
  • FIG 4 illustrates a side view of device 300 of FIG 3 for culturing a plurality of biological samples, according to some embodiments of the invention.
  • the figure offers a view of inlet 303, from where fluids enter the device.
  • the fluids flow in the annular fluid path, until they reach the splits, from each split the fluids are pushed via an orifice 401 into the respective carrier 302, where the first portion receives them.
  • the level of the first portion rises until reaching an opening 402 in the culturing module's side wall, from where the out-flow flows to the hollow space 403 of the chamber, as represented by an arrow enumerated 404.
  • the received flow 404 of culturing fluids form part of the second portion.
  • the devices illustrated provide closed systems, that is, the devices are configured to isolate the culturing fluid and the biological sample from direct contact with an environment external to the device. This way, if a device is kept sterile until after the sample is inserted thereto and the device is closed (e.g., by positioning the carrier in FIG 1, or by closing the lid in FIG 2A, or by any other mean applicable to the case), a user of the device may take it into a non- sterile environment, while the culturing fluid and the biological sample enclosed therein remain sterile.
  • FIG 5 schematically illustrates embodiments of the invention wherein a pump is used to drive flow. Due to convenience considerations, the embodiments are schematically illustrated as resembling the embodiments of FIG 2B, however this is non-limiting and the description provided is applicable for any other embodiment, unless specifically noted otherwise.
  • FIG 5 depicts a device 500 comprising a culturing module 501 and a chamber 502, coupled via a fluidic connection 503, which is a conduit in this case.
  • a circulator 504 is depicted, while the circulator is also a conduit.
  • the two illustrated conduits503 and 504 are configured to drive flow of fluids in device 500, by an external pump 505 and 506, respectively.
  • the external pump may be, for example, a peristaltic pump, as illustrated in FIG 5A, although it is non-limited thereto.
  • the culturing modules described so far, and possibly additional or alternative embodiments of culturing modules are configured to hold a culturing fluid.
  • they are containers configured to hold a culturing fluid (i.e., the first portion) and configured to allow culturing a biological sample in the first portion.
  • the chambers are also containers, in turn configured to hold a second portion of the culturing fluid.
  • the embodiments schematically illustrated in, e.g., FIGs 1, 2A and 2B and others each comprises two containers.
  • the devices disclosed by embodiments of the preset invention are devices for culturing biological samples, such as embryos. It is known in the art that tissue culturing should be done in a temperature substantially equal to a physiological temperature of the culture's source. For example, in human the physiological temperature is substantially equal to 37°C, while in sheep it is substantially 38.5°C. It is appreciated that in mammals, for example, the physiological temperature is higher than normal room temperature, and therefore it is required to warm the environment around the biological sample to the proper temperature.
  • the sample is submerged within a culturing fluid, it is appreciated that warming the fluid substantially to a physiological temperature will provide the sample with an environment whose temperature is proper for the culturing thereof.
  • the fluid's temperature should be substantially at a physiological temperature. It is known in the art that the temperature may deviate slightly from the expected physiological temperature within a lower limit, below which the sample is damaged, and a higher limit, above which the sample is damaged as well. Hence, the temperature of the fluid should be between the lower limit and the upper limit.
  • embodiments of the present invention that allow warming the fluids within the device may have heat conducting walls, at least in some elements of the device. It is then possible to place the device in a warm room, or chamber, wherein the temperature is as required.
  • the bottom, lower part of the culturing module may be heat conducting. Therefore, it is possible to place the culturing module alone (instead of placing the whole device), e.g., in a warm bath, thereby warming the fluids in the first portion, in which the sample is submerged.
  • yet other embodiments may comprise a chamber whose bottom walls are heat conducting. Therefore, placing the chamber in a warm environment, such as a warm bath, will warm the fluid in the second portion, while flow should bring the warm fluid to the culturing module and to the sample.
  • the fluid remains warm through most of the time. Flow from the chamber to the culturing module is not immediate. Therefore the fluid should be warmed in the chamber probably to a temperature higher than the required physiological temperature, so as to reach the culturing module in the required temperature. While in the culturing device the fluid is expected to be at a physiological temperature, it can be appreciated that upon returning to the chamber the fluid's temperature will still be higher than room temperature.
  • FIG 6 schematically illustrates a device 600 comprising a chamber 601 and a culturing module 602 coupled via a fluidic path, according to some embodiments of the invention.
  • the device has a circulator 604.
  • the bottom of the chamber is placed in a cooling unit 605.
  • the fluid at the culturing module would be cold.
  • FIG 7 schematically illustrates a device 700 for culturing a biological sample configured to submerge the sample in a warm culturing fluid.
  • device 700 resembles device 600 of FIG 6, and the bottom of the chamber thereof is being placed in a cooling unit.
  • device 700 comprises a heating container 701 on the circulation path of the culturing fluid.
  • the heating container is placed between the chamber and the culturing module and is couple thereto via a conduit 703 and conduit 704 respectively.
  • the heating container is placed in a heating unit 702, such as a warm bath, a closed box heated by air etc. Fluid from the second portion flows via conduit 703 to heating container 701, configured to hold a third portion of culturing fluid.
  • the culturing fluid in the third portion is warmed to above the physiological temperature, and then flows to the culturing device.
  • FIG 8 provides another device 800 for culturing a biological sample configured to submerge the sample in a warm culturing fluid, according to some embodiments of the invention.
  • the conduit of the circulator is "replaced" by a long conduit 802 coiling around a heating unit 803.
  • Heating unit 803 may be a heating element as illustrated in the figure, although it may be any other heating unit applicable to the case, such as a cylinder containing warm liquid etc.
  • the culturing fluid flows in the coiled conduit 803 until it is warmed to a temperature warmer than the desired physiological temperature, whereupon it flows to the culturing module 804. Before the fluid reaches culturing module 804 it slightly colds down, therefore reaching culturing module 804 in the desired temperature.
  • thermocouple 805 coupled to pump 806, in the flow-path, before the fluid reaches culturing module 804, thereby assuring the fluid would not enter culturing module 804 too warm, thus damaging the sample. If the first thermocouple 805 detects that the fluid it too warm it may be configured to turn the pump off, until the fluid colds down, hence preventing damage to the sample.
  • FIG 8 the pump in FIG 8 is illustrated between the chamber and the coil. However, this is non-limiting and any other embodiment applicable to the case may be used, s explained, for example, with reference to FIG 5.
  • thermocouple 807 near or after the fluid leaves the coil on its way to culturing module 804. Second thermocouple 807 may also be coupled to pump 806. If second thermocouple 807 detects that the fluid is too warm or too cold while leaving the coil, it may instruct pump 806 to speed up or to speed down the flow, hence causing the fluid to stay shorter or longer, accordingly, in the coil and around the heating unit 803, thereby affecting its temperature.
  • the fluid's temperature should be substantially at a physiological temperature. It is known in the art that the temperature may deviate slightly from the expected physiological temperature within a lower limit, below which the sample is damaged, and a higher limit, above which the sample is damaged as well. Hence, the temperature of the fluid should be between the lower limit and the upper limit.
  • FIG. 9 schematically illustrates a device 902 for culturing a biological sample, according to some embodiments of the invention.
  • Device 902 is coupled via a conduit 904 or via any other mean applicable to the case to the source container 901.
  • the fluid flows from source container 901 to a waste chamber 903, coupled to device 902, e.g., via conduit 905.
  • the waste chamber may be or may comprise, for example, the chamber described above. This way it may be possible to continuously provide the device with fresh culturing fluid instead of with circulated fluid.
  • An external pump 906 can be coupled, e.g., to conduit 904, to the outlet of source container 904, or at any other applicable position.
  • the coil e.g., along conduit 904 in order to warm the fluid, e.g., as was previously presented in FIG 8. It should be mentioned, though, that in the illustrated embodiments of FIG. 9, the volume of the hollow space of the waste chamber should be large enough to store al the culturing fluid provided throughout the culturing of the sample.
  • fresh fluids are provided at a rate of, e.g., 1 milliliter per hour, within 24 hours there will be 24 milliliters of culturing fluid in the chamber.
  • Culturing an embryo may take four days, for example, and therefore the chamber should be able to store approximately 100 milliliters of fluids.
  • the device facilitates continuous flow of culturing fluid inside the culturing chamber, as previously illustrated as explained with reference to the embodiments described above.
  • outlet 907 of device 902 may be coupled directly or indirectly (e.g., via conduit 905) to drainage means for disposing the fluid (such as sewage), or waste chamber 903 may allow emptying accumulated fluid, etc.
  • Device 902 may be or may comprise, for example, a culturing module as explained previously with reference to any of the embodiments mentioned above. Moreover, device 902 may be stored in a temperature controlled chamber 908, thus providing a warm environment that supports culturing, e.g., en environment where the temperature is a physiological temperature. If the device is operated in an environment warmer than the physiological temperature, temperature controlled chamber 908 may cold the environment around the culturing chamber instead of warming it. In some embodiments, source container 901 may be stored in a cold chamber 909, such as a refrigerator, for keeping the source culturing fluid refrigerated, thus preventing contamination. Alternatively, it should be appreciated that in some cases the source container may be replaced from time to time with a fresh container, as an additional or alternative way for preventing contamination.
  • a cold chamber 909 such as a refrigerator

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Thermal Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Molecular Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention concerne un dispositif pour la culture d'un échantillon biologique dans un fluide de culture. Le dispositif comprend un module de culture. Le module de culture est configuré pour maintenir une première partie de la culture fluide en flux continu et pour permettre la culture de l'échantillon biologique dans la première partie, le liquide de culture dans la première partie étant essentiellement à une température physiologique de l'échantillon biologique. Le dispositif peut également comprendre une chambre, configurée de façon à maintenir une seconde partie du fluide de culture dans un espace creux de la chambre, le liquide de culture dans la seconde partie étant plus froid que la température du fluide dans la première partie.
PCT/IL2015/050698 2014-07-06 2015-07-06 Dispositifs pour la culture d'un échantillon biologique dans un liquide de culture WO2016005971A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ILPCT/IL2014/050608 2014-07-06
IL2014050608 2014-07-06

Publications (1)

Publication Number Publication Date
WO2016005971A1 true WO2016005971A1 (fr) 2016-01-14

Family

ID=55063677

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IL2015/050698 WO2016005971A1 (fr) 2014-07-06 2015-07-06 Dispositifs pour la culture d'un échantillon biologique dans un liquide de culture

Country Status (1)

Country Link
WO (1) WO2016005971A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996040858A1 (fr) * 1995-06-07 1996-12-19 Aastrom Biosciences, Inc. Appareil et procede destines a la conservation et a la culture de cellules biologiques
US5846828A (en) * 1995-06-07 1998-12-08 Advanced Tissue Sciences Apparatus and method for sterilizing, seeding, culturing, storing, shipping, and testing tissue, synthetic, or mechanical heart valves orvalve segments
WO2009039433A1 (fr) * 2007-09-20 2009-03-26 Incept Biosystems Inc. Système de culture analytique microfluidique

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996040858A1 (fr) * 1995-06-07 1996-12-19 Aastrom Biosciences, Inc. Appareil et procede destines a la conservation et a la culture de cellules biologiques
US5846828A (en) * 1995-06-07 1998-12-08 Advanced Tissue Sciences Apparatus and method for sterilizing, seeding, culturing, storing, shipping, and testing tissue, synthetic, or mechanical heart valves orvalve segments
WO2009039433A1 (fr) * 2007-09-20 2009-03-26 Incept Biosystems Inc. Système de culture analytique microfluidique

Similar Documents

Publication Publication Date Title
RU2371375C2 (ru) Разливной автомат для напитка
US20100209966A1 (en) Aseptic bioreactor system for processing biological materials
CN102006807B (zh) 对诸如瓶子和注射器等母乳容器的加温特别有用的母乳处理装置
CN113227350B (zh) 具有受控环境区的细胞培养和组织工程系统
RU2014127153A (ru) Терморегулируемые несущие поверхности для гибких настенных систем одноразового использования
RU2005128289A (ru) Система контейнера для инкубации и/или хранения и способ
CN107557297A (zh) 用于生物材料的联合式培育和玻璃化的设备
JP2007508025A (ja) 二つのキャビネットを備えた実験室用装置
KR101224110B1 (ko) 딸기 육묘용 장치
JP5072039B2 (ja) 飲料サーバー
WO2016005971A1 (fr) Dispositifs pour la culture d'un échantillon biologique dans un liquide de culture
ES2856854T3 (es) Unidad de refrigeración
KR20220084779A (ko) 냉난방이 가능한 수경 재배 시스템
CN201648374U (zh) 一种微生物培养箱
KR101534114B1 (ko) 배양기
JP4708995B2 (ja) 観賞魚展示販売用多段型水槽台
CN215713047U (zh) 一种灭菌型细胞培养箱
US20220001380A1 (en) Microscope for microscopic examination of a sample
BE1017992A3 (nl) Bewaarkast.
EP3011827A1 (fr) Système de climatisation de serres
KR102604194B1 (ko) 세포·미생물 배양용기장치의 순환수 제어 시스템 및 제어방법
CN210176844U (zh) 一种改进的生化培养箱
JP2017063695A (ja) 株元温度制御機構付栽培装置
AU2023378831A1 (en) A modular incubator system
JP6551504B2 (ja) 培地供給装置

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15818330

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15818330

Country of ref document: EP

Kind code of ref document: A1

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载