US 12,441,983 B2
Suspension culture of human embryonic stem cells
Ramkumar Mandalam, Union City, CA (US); Yan Li, Tallahassee, FL (US); and Isabelle Nadeau-Demers, San Francisco, CA (US)
Assigned to Asterias Biotherapeutics, Inc., Carlsbad, CA (US)
Filed by ASTERIAS BIOTHERAPEUTICS, INC., Carlsbad, CA (US)
Filed on Jan. 24, 2024, as Appl. No. 18/421,847.
Application 18/421,847 is a continuation of application No. 16/862,559, filed on Apr. 29, 2020.
Application 16/862,559 is a continuation of application No. 15/894,842, filed on Feb. 12, 2018, granted, now 10,676,714, issued on Jun. 9, 2020.
Application 15/894,842 is a continuation of application No. 14/791,479, filed on Jul. 6, 2015, abandoned.
Application 14/791,479 is a continuation of application No. 11/917,993, granted, now 9,074,181, issued on Jul. 7, 2015, previously published as PCT/US2006/023976, filed on Jun. 20, 2006.
Claims priority of provisional application 60/693,266, filed on Jun. 22, 2005.
Prior Publication US 2024/0158741 A1, May 16, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/0735 (2010.01)
CPC C12N 5/0606 (2013.01) [C12N 2501/115 (2013.01); C12N 2501/125 (2013.01); C12N 2501/15 (2013.01); C12N 2501/26 (2013.01)] 21 Claims
 
1. A method of maintaining human pluripotent stem (hPS) cells in an undifferentiated state comprising:
a) suspending human pluripotent stem (hPS) cells in a nutrient medium to create a suspension culture, wherein the nutrient medium comprises 20 ng/mL to 500 ng/mL of fibroblast growth factor (FGF) and transforming growth factor beta (TGFβ), wherein the nutrient medium does not contain serum;
b) culturing the hPS cells in the suspension culture for a period of time; and
c) adding fresh nutrient medium to the suspension culture, wherein the fresh nutrient medium does not contain serum; wherein the hPS cells are not grown on a solid substrate.