Media and methods for enhancing the survival and proliferation of stem cells
The present disclosure relates to improved supplements, culture media and methods for enhancing the survival or proliferation of mammalian stem cells. In particular, adding a lipid supplement, such as a lipid-enriched carrier (e.g. a lipid-enriched albumin), to the culture medium may enhance the survival and/or proliferation of the stem cells by at least 5% to 65% as compared to a culture medium that does not contain the lipid supplement.
1. A cloning supplement comprising one or more small molecule inhibitors and two or more lipids in the presence of a carrier, wherein
the carrier is an agent, substance, composition or complex that transports some or all of the two or more lipids to a mammalian pluripotent stem cell,
the two or more lipids comprise oleic acid and palmitic acid,
the amount of each alpha-linolenic acid and/or arachidonic acid is less than 300 μg per gram of carrier, and
wherein the survival or proliferation of mammalian pluripotent stem cells is enhanced in a culture medium containing the supplement as compared to a culture medium that does not contain the supplement.
2. The cloning supplement according to claim 1 , further comprising one or more of Mead's acid, arachidic acid, palmitoleic acid, myristic acid, myristoleic acid, linoleic acid, stearic acid, alpha-linolenic acid, arachidonic acid, cholesterol, and DL-alpha-tocopheryl.
3. The cloning supplement according to claim 1 , wherein the cloning efficiency is 5% to 65%.
4. The cloning supplement according to claim 1 , wherein the pluripotent stem cells are ES cells or iPS cells.
5. The cloning supplement according to claim 1 , wherein the carrier is an agent for transporting the one or more lipids to the one or more mammalian pluripotent stem cells.
6. The cloning supplement according to claim 1 , wherein the carrier is an albumin, a liposome, an extracellular vesicle, an exosome, a nanostructured lipid carrier, or a cyclodextrin.
7. The cloning supplement according to claim 6 , wherein the albumin is human.
8. The cloning supplement according to claim 6 , wherein the albumin is recombinant.
9. The cloning supplement according to claim 1 , wherein at least some of the one or more lipids are bound to the carrier.
10. The cloning supplement according to claim 1 , wherein a concentration of the one or more lipids ranges from 1 ng/ml to 35 μg/ml.
11. The cloning supplement according to claim 1 , wherein the cloning supplement is diluted in the mammalian pluripotent stem cell culture medium.
12. The cloning supplement according to claim 1 , wherein the one or more small molecule inhibitors is Thiazovivin, Y-27632, CHIR99021, S8202190, MI-7, Necrostatin-1, NS3694, Wnt-C59, NSCI, or BIPV5.