What is your preferred method for confirming pluripotency of iPSCs from blood cells?

Answer

Pluripotency can only be confirmed by tri-lineage differentiation either through in vivo formation of a teratoma containing all three germ layers or by in vitro differentiation to cells of each germ layer.

Our preferred method would be in vitro directed differentiation using:

STEMdiff™ Definitive Endoderm kit for differentiation,

STEMdiff™ NIM for neural differentiation, and

One of many STEMdiff™ APEL™-based protocols.

Expression of markers associated with the undifferentiated state, including expression of Oct3/4, Sox2, Tra-1-60, Tra-1-81, SSEA3 and SSEA4, should be considered a screening tool and not a replacement for a tri-lineage differentiation assay.