Characterization of CD30 expression in human embryonic stem cell lines cultured in serum-free media and passaged mechanically

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Abstract
The presence of chromosomal abnormalities could have a negative impact for human embryonic stem cell (hESC) applications both in regenerative medicine and in research. A biomarker that allows the identification of chromosomal abnormalities induced in hESC in culture before they take over the culture would represent an important tool for defining optimal culture conditions for hESC. Here we investigate the expression of CD30, reported to be a biomarker of hESCs with abnormal karyotype, in undifferentiated and spontaneously differentiated hESC. hESC were derived and cultured on mouse fibroblasts in KO–SR containing medium (serum free media) and passaged mechanically. Our results based on analysis at mRNA (RT–PCR) and protein (fluorescence-activated cell sorting and immunocytochemistry) level show that CD30 is expressed in undifferentiated hESC, even at very early passages, without any correlation with the presence of chromosomal anomalies. We also show that the expression of CD30 is rapidly lost during early spontaneous differentiation of hESC. We conclude that CD30 expression in hESC cultures is probably a consequence of culture conditions, and that KO–SR may play a role. In addition, the expression of so-called ‘stemness’ markers does not change in undifferentiated hESC during long-term culture or when cells acquire chromosomal abnormalities.

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