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4C,D). The maximal decrease of proliferation was 52.9?��?3.35% for ASCs and 81.24?��?4.40% for BM-MSCs, respectively, at 1250?ng/mL. ASCs had higher levels of proliferation on day 1 with gradual reduction in proliferation toward day 7 (Figure ?(Figure4C),4C), while BM-MSC proliferation improved with tacrolimus incubation reaching higher levels at day 7 (Figure ?(Figure44D). Figure 4 Cell proliferation under drug exposure. CyQUANT assay results with a clear dose-dependent effect for ASCs and BM-MSCs under ALS-influence on days 1 (A) and 7 (B). CyQUANT assay results after adding tacrolimus on day 1 (C) and day 7 (D). CyQUANT assay ... Serum effects on cell viability Replacing the culture medium of stem cells in vitro with serum from ALS-treated rats reduced the viability of ASCs significantly (Figure ?(Figure5A).5A). On day 1, viability decreased to 71.53?��?21.27%, which was significant compared to day 2. Tests performed at later time points did not reveal Selumetinib this early detrimental effect. BM-MSCs, however, showed a similar but delayed decrease of viability with significant differences after day 2 (Figure ?(Figure55B). Figure 5 ASC and BM-MSC susceptibility to ALS. ASC [blue bars, (A)] and BM-MSC [red bars, (B)] were incubated for 2?h in 100% serum from ALS-treated or untreated na?ve rats. An MTT assay was performed on days 1, 2, 4, and 8. The direct absorbance ... Complement-dependent cytotoxicity assay The CDC assay was utilized to detect cell lysis based on glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-release SP600125 cost using rat complement as a probe in the presence of ALS. Whereas the negative control (cells and 40?��L/mL of ALS) did not show a strong response (Figure Thymidine kinase ?(Figure6),6), low concentrations of ALS revealed a CDC with a strong dose-dependency. ASCs did not succumb to these complement-mediated effects to the same extent as BM-MSCs and showed lower levels of luminescence compared to that with BM-MSCs (Figure ?(Figure6A6A vs. Figure ?Figure6B).6B). Maximal lysis control was obtained by the combination of cells, complement, ALS (40?��L/mL), and lysis buffer. Figure 6 Complement-dependent cytotoxicity (CDC) under drug exposure. CDC assay results for ASCs (A) and BM-MSCs (B). Higher concentrations of ALS showed cytotoxic effects on BM-MSCs but not ASCs. *P?