US 11,959,098 B2
Methods and culture media for culturing pluripotent stem cells
Michal Amit, Yuvalim (IL); and Joseph Itskovitz-Eldor, Haifa (IL)
Assigned to Technion Research & Development Foundation Limited, Haifa (IL)
Filed by Technion Research & Development Foundation Limited, Haifa (IL)
Filed on Mar. 5, 2020, as Appl. No. 16/809,771.
Application 16/809,771 is a continuation of application No. 15/403,303, filed on Jan. 11, 2017, granted, now 10,597,635, issued on Mar. 24, 2020.
Application 15/403,303 is a continuation of application No. 13/821,244, granted, now 10,214,722, issued on Feb. 26, 2019, previously published as PCT/IL2011/000722, filed on Sep. 7, 2011.
Claims priority of provisional application 61/380,388, filed on Sep. 7, 2010.
Prior Publication US 2020/0224157 A1, Jul. 16, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/074 (2010.01); C12N 5/0735 (2010.01); C12N 5/077 (2010.01); C12N 5/0775 (2010.01); C12N 5/079 (2010.01)
CPC C12N 5/0607 (2013.01) [C12N 5/0606 (2013.01); C12N 5/0653 (2013.01); C12N 5/0654 (2013.01); C12N 5/0655 (2013.01); C12N 5/0696 (2013.01); C12N 5/0618 (2013.01); C12N 5/0662 (2013.01); C12N 2500/90 (2013.01); C12N 2500/92 (2013.01); C12N 2500/98 (2013.01); C12N 2501/115 (2013.01); C12N 2501/13 (2013.01); C12N 2501/23 (2013.01); C12N 2501/2306 (2013.01); C12N 2501/2311 (2013.01); C12N 2501/237 (2013.01); C12N 2506/02 (2013.01); C12N 2506/03 (2013.01); C12N 2506/45 (2013.01); C12N 2509/10 (2013.01)] 13 Claims
 
1. A method of expanding and maintaining pluripotent stem cells (PSCs) in an undifferentiated state, the method comprising:
(a) passaging the PSCs in a suspension culture by mechanical dissociation of PSC clumps to single cells or clusters under conditions devoid of an enzymatic dissociation, each of said clusters comprising no more than 200 PSCs, for at least 2 and no more than 10 passages, to thereby obtain a suspension culture of PSCs devoid of clumps; and
(b) passaging without dissociation said suspension culture of PSCs devoid of said clumps and comprising single cells or small clusters, each of the clusters comprising no more than 200 PSCs,
thereby expanding and maintaining the PSCs in the undifferentiated state.