Human UCB-MSCs show expression of pluripotent and neural progenitor markers. (A) RT-PCR analysis shows the presence of transcripts corresponding to pluripotent as well as neural progenitor markers in freshly isolated human umbilical cord blood-derived (hUCB) mononuclear cell MNC01 and also in expanded mesenchymal stem cell (hUCB-MSC) SCB4 (lanes 4 and 5). hUCB MNC01 showed a significant level of expression of pluripotent markers but lacked nestin expression when compared with control RCB 2080 (lane 2), whereas hUCB-MSC SCB4 showed a higher level of nestin and reduced level of pluripotent marker expression. (B) Comparison of the relative pixel intensity of gene expression in hUCB MNC01. (C) Comparison of the relative pixel intensity of gene expression in RCB 2080 and hUCB-MSC SCB4. (D) Comparison of pluripotent marker and neural stem cell marker expression in two more sets of hUCB-MNCs (MNC02 and MNC03) and hUCB-MSCs (hUCB-MSC SCB10 and SCB14), which also shows a varying pattern of expression corroborating our previous observations that neural stem cell marker expression varies from sample to sample. (E) Relative pixel intensity of gene expression in hUCB MNC02 and MNC03. (F) Relative pixel intensity of gene expression in hUCB-MSC SCB10 and hUCB-MSC SCB14. (G) to (R) Treatment of control RCB 2080, hUCB-MSC SCB4 and hUCB-MSC SCB10 cells with neurobasal (NB) medium in combination with and without 0.5 μm retinoic acid (RA), 1% FBS and 10 ng basic fibroblast growth factor (bFGF). Scale bar = 100 μm.