Expression of CD146 and Regenerative Cytokines by Human Placenta-Derived Mesenchymal Stromal Cells upon Expansion in Different GMP-Compliant Media

Figure 3

Time line of PLL effects on the expression of CDC146. The pMSC from 4 donors was expanded in LM to their second passage and further expanded in LM+5% (vol/vol) PLL. Controls were incubated in LM. (a) The expression of CD146 on cells surfaces was investigated by flow cytometry. Seven days after addition of PLL, the MFI of the CD146 expression dropped significantly in comparison to pMSCs in LM and in comparison to cells 1 day after addition of PLL ( each). (b) The expression of the total CD146 protein was investigated by immunoblot. Seven days after addition of the PLL total protein of the CD146 expression dropped significantly in comparison to pMSCs in LM and in comparison to cells 1 day after addition of PLL ( each). (B’) A representative immunoblot is shown in the lower right: CD146 is not detected in control cells (lane 1), but in pMSCs expanded for 7 days in LM (lane 2), almost not in pMSCs expanded for 7 days in LM + PLL (lane 3). CD146 is detected in pMSCs expanded for 72 hours (lane 4) or 24 hours (lane 5) in LM + PLL. Detection of ß-actin served as loading control. Size markers are shown in lanes M. The molecular weight of proteins separated by SDS-PAGE is noted on each blot. (c) The expression of transcripts encoding CD146 was enumerated by RT-PCR. After 24 hours of incubation in LM + PLL, a significant drop in CD146 transcripts was noted in comparison to pMSCs in LM (). After 3 and 7 days, CD146 encoding transcript increased somewhat but stayed well below the levels recorded in pMSCs in LM.

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