Efficient derivation of stable pluripotent bovine embryonic stem cells
AssigneesThe Regents of the University of California, The Salk Institute for Biological Studies
InventorsPablo Juan Ross, Yanina Bogliotti, Marcela Vilarino, Juan Carlos Izpisua Belmonte, Jun Wu
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Abstract
This disclosure provides ungulate embryonic stem cells (ESCs) derived from the inner cell mass of pre-implantation blastocysts or pluripotent cells from embryos. From an agricultural and biomedical perspectives, the derivation of stable ESCs from domestic ungulates is important for genomic testing and selection, genetic engineering, and providing an experimental tool for studying human diseases. Cattle are one of the most important domestic ungulates that are commonly used for food and bioreactors.
Claims9
A method for culturing pluripotent bovine embryonic stem cells (ESCs), comprising culturing a bovine ESC in the presence of extracellular matrix components produced by Englebreth-Holm-Swarm sarcoma cells or Vitronectin, without inactivated feeder cells, in a cell culture media comprising:
- (i) an effective amount of Activin-A;
- (ii) an effective amount of Fibroblast Growth Factor 2 (FGF2);
- and (i ii) an effective amount of one or more inhibitors of canonical Wnt signaling,
- thereby preparing cultured bovine ESCs;
- wherein the cultured bovine ESCs are stable bovine ESCs that express stable morphology, karyotype and/or pluripotent marker expression and remain stable for at least 34 passages.
Dependencies are inferred from the claim text.