Micromanipulation and storage apparatus and methods

• US 10,244,749 B2
• Filed: 06/24/2016
• Issued: 04/02/2019
• Est. Priority Date: 05/28/2010
• Status: Active Grant

• Current Assignee: Genea Ip Holdings Pty Limited
• Original Assignee: Genea Ip Holdings Pty Limited
• Inventors: Henderson, Chester John, Lewis, Craig Matthew, Roy, Tammie Kim, Vom, Eduardo
• Primary Examiner(s): Barron, Sean C.
• Application Number:
  US15/192,349
• Publication Number:
  US 20160316743A1
• Time in Patent Office: 1,012 Days
• CPC Class Codes: A01N 1/0263 Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
• CPC Class Codes: A01N 1/0268 Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen
• CPC Class Codes: A01N 1/0284 Temperature processes, i.e. using a designated change in temperature over time
• CPC Class Codes: A61B 17/43 for artificial insemination
• CPC Class Codes: A61B 17/435 for embryo or ova transplantation
• CPC Class Codes: A61D 19/022 Containers for animal semen, e.g. pouches or vials A61D19/027 takes precedence; Methods or apparatus for treating or handling animal semen containers, e.g. filling or closing
• CPC Class Codes: A61D 19/025 with means for controlling the temperature, e.g. heating or cooling
• CPC Class Codes: A61D 19/04 for embryo transplantation

Abstract

A method of cryopreservation of biological material includes the steps of providing an apparatus including a vessel having a reservoir wherein the vessel has a channel formed in a portion of the reservoir, the channel including an intermediate restriction dimensioned to resist passage of the biological material but allow passage of liquid treatment solutions, and the channel including sub-reservoirs at either end of the restriction; introducing the biological material into a reservoir region of the apparatus for capture in a channel region of the vessel; irrigating and draining the biological material with a series of vitrification solutions introduced into a first of the sub-reservoirs and removed from a second of the sub-reservoirs; final draining the irrigation solutions from the vessel; and vitrifying the vessel and captive biological material.

Claims

1. A method of cryopreservation of biological material using an apparatus including a vessel having a reservoir, wherein said vessel has a channel formed in a portion of said reservoir, said channel including an intermediate restriction dimensioned to resist passage of said biological material but allow passage of liquid treatment solutions, and said channel including a first sub-reservoir at a first end of said restriction and a second sub-reservoir at a second end of said restriction, the method comprisin the steps of:

introducing said biological material into the first sub-reservoir of said vessel;

irrigating said biological material with a series of vitrification solutions introduced into the first sub-reservoir and removed from the second sub-reservoir;

removing the vitrification solutions from the second sub-reservoir, leaving a residual volume of liquid surrounding the biological material that is sufficiently small for vitrification; and

vitrifying the biological material and the residual volume of liquid surrounding the biological material within the vessel.

2. A method according to claim 1, wherein said vitrification solutions include non-permeating and permeating cryoprotectants selected from the group consisting of Ethylene Glycol (EG), Dimethyl Sulphoxide (DMSO), Glycerol, and disaccharides solutions.

8. A method according to claim 1, wherein said vitrification solutions are gradually introduced at a rate to enhance sinking of said biological material to a bottom of the first sub-reservoir.

9. A method according to claim 1, wherein said vitrification solutions include non-permeating cryoprotectants selected from disaccharides solutions.

11. A method according to claim 1, wherein said vitrification solutions include permeating cryoprotectants selected from Ethylene Glycol (EG), Dimethyl Sulphoxide (DMSO), and Glycerol.

3. A method of cryopreservation of biological material using an apparatus including a vessel having a reservoir, wherein said vessel has a channel formed in a portion of said reservoir, said channel including an intermediate restriction dimensioned to resist passage of said biological material but allow passage of liquid treatment solutions, and said channel including a first sub-reservoir at a first end of said restriction and a second sub-reservoir at a second end of said restriction, the method comprising the steps of:

introducing said biological material into the first sub-reservoir of said vessel;

irrigating said biological material with a single vitrification solution gradually introduced into the first sub-reservoir;

removing the vitrification solution from the second sub-reservoir, leaving a residual volume of liquid surrounding the biological material that is sufficiently small for vitrification;

vitrifying the biological material and the residual volume of liquid surrounding the biological material within the vessel.

4. A method according to claim 3, wherein said vitrification solution includes non-permeating and permeating cryoprotectants selected from the group consisting of Ethylene Glycol (EG), Dimethyl Sulphoxide (DMSO), Glycerol, and disaccharides solutions.

5. A method according to claim 3, wherein said vitrification solution is gradually introduced at a rate to enhance sinking of said biological material to a bottom of the first sub-reservoir.

10. A method according to claim 3, wherein said vitrification solution includes non-permeating cryoprotectants selected from disaccharides solutions.

12. A method according to claim 3, wherein said vitrification solution includes permeating cryoprotectants selected from Ethylene Glycol (EG), Dimethyl Sulphoxide (DMSO), and Glycerol.

6. A method for pre-vitrification treatment of a biological material, using an apparatus including a vessel having a reservoir wherein said vessel has a channel formed in a portion of said reservoir, said channel including an intermediate restriction dimensioned to resist passage of said biological material but allow passage of liquid treatment solutions and said channel including a first sub-reservoir at a first end of said restriction and a second sub-reservoir at a second end of said restriction, the method comprising the steps of:

introducing said biological material into the first sub-reservoir of said vessel; and

irrigating said biological material with pre-vitrification solutions introduced into the first sub-reservoir and removed from the second sub-reservoir.

7. A method of thawing a vitrified biological material in an apparatus including a vessel having a reservoir wherein said vessel has a channel formed in a portion of said reservoir, said channel including an intermediate restriction dimensioned to resist passage of said biological material but allow passage of liquid treatment solutions, and said channel including a first sub-reservoir at a first end of said restriction and a second sub-reservoir at a second end of said restriction, the method comprising the steps of:

heating the vessel;

irrigating the biological material with a series of thawing solutions introduced into the first sub-reservoir and removed from the second sub-reservoir; and

recovering the thawed biological material from said vessel.