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Fig. 3 | Stem Cell Research & Therapy

Fig. 3

From: A robust potency assay highlights significant donor variation of human mesenchymal stem/progenitor cell immune modulatory capacity and extended radio-resistance

Fig. 3

Pooled pre-labeled PBMC and pooled reference MSPC make a robust assay format to readout inhibition of T-cell proliferation in an off-the-shelf potency assay. a, b Pooled carboxyfluorescein pre-labeled random donor peripheral blood mononuclear cell (pPBMC) aliquots seeded in triplicate show a highly significant mitogen-induced proliferation (phytohemagglutinin (PHA); green bars) compared to minimum proliferation of the unstimulated pPBMC seeded off-the-shelf in the absence of PHA (dark grey bar) at day 4 (d4). Triplicates of pMSPC composed of cells from five each random donors (D1D15) of bone marrow (BM; blue bars), white adipose tissue (WAT; yellow bars) and umbilical cord (UC; red bars) were used as an organotypic MSPC reference (grey areas) to determine organ-specific highly significant inhibition of mitogen-induced T-cell proliferation compared to individual a cultured or b freshly thawed individual MSPC from five donors per organ. c, d At day 7 (d7), the potent allo-response of pPBMC (grey bars; for time course titration see Fig. 1b, c) in the absence of external stimulation was significantly inhibited by some but not all individual MSPCs compared to the organotypic reference pMSPC (grey areas; same donors and identical color code as in a, b). Differences between c cultured and d freshly thawed individual or pooled MSPCs were more prominent compared to the inhibition of mitogenesis (in a, b). Significant inhibition of day 4 mitogen-induced (a, b) and day 7 mixed leukocyte reaction (MLR)-induced (c, d) pooled T-cell proliferation is indicated at the right margin of the graphs; significance of individual donor comparison is indicated by vertical lines. Mean ± SD results at an MSPC:PBMC ratio of 1:3 are shown. *p < 0.05; **p < 0.01 ***p < 0.001; ****p < 0.0001

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