US 12,167,729 B2
Cryopreservation of tissues and organs
Mustafa Korkut Uygun, Newton, MA (US); Shannon Noella Tessier, Framingham, MA (US); and Renier Johan de Vries, Boston, MA (US)
Assigned to The General Hospital Corporation, Boston, MA (US)
Appl. No. 17/058,279
Filed by The General Hospital Corporation, Boston, MA (US)
PCT Filed May 30, 2019, PCT No. PCT/US2019/034744
§ 371(c)(1), (2) Date Nov. 24, 2020,
PCT Pub. No. WO2019/232268, PCT Pub. Date Dec. 5, 2019.
Claims priority of provisional application 62/677,999, filed on May 3, 2018.
Prior Publication US 2021/0195891 A1, Jul. 1, 2021
Int. Cl. A01N 1/02 (2006.01)
CPC A01N 1/0284 (2013.01) [A01N 1/0221 (2013.01); A01N 1/0247 (2013.01)] 29 Claims
 
1. A method for preserving a biological sample, the method comprising:
(a) perfusing the biological sample at a first perfusion rate with a first loading solution comprising one or more cryoprotective agent(s) at a hypothermic temperature between 0° C. and 12° C.;
(b) perfusing the biological sample at a second perfusion rate lower than the first perfusion rate with a second loading solution comprising a higher concentration of at least one of the one or more cryoprotective agent(s) than in the first loading solution at a hypothermic temperature;
(c) placing the perfused biological sample in a container;
(d) removing air from the container to avoid ice crystal formation in cells within the biological sample or the first and second loading solutions, and sealing the container; and
(e) cooling the biological sample in the container to a subzero temperature, thereby preserving the biological sample at a subzero temperature.