Fig. 1
BM-MSC support in vitro survival of blood ASC. a Short-term survival of blood ASC in irradiated MSC (iMSC) co-cultures. FACS purified blood ASC (1000/well) were cultured in RPMI with 10% FBS (R10) (No MSC) or co-cultured with 50,000 BM-MSC (MSC) for 7 days (p < 0.001; ANOVA). Representative images of Elispot wells are shown. b Long-term survival of blood ASC and iMSC co-cultures from two different adult blood samples after vaccination. Five hundred to 1500 ASC with 25,000 or 30,000 iMSC were co-cultured in each well and IgG Elispots were performed on designated days. c, d Pro-survival support of BM-MSC is independent of cell–cell contact. Same number of blood ASC were cultured in R10 (Media), co-cultured with 50,000 iMSC (iMSC), in transwells with ASC and 50,000 iMSC in separate chambers (TW), or iMSC secretome (Secretome). Representative Elispot wells are shown in d. e Long-term survival of blood ASC in MSC secretome. One thousand ASC were cultured in MSC secretome or with media alone (p < 0.03; linear regression modeling using media (secretome) as a covariate). In a–c, e, Elispot assays were performed on the indicated day and the frequency (%) of IgG-secreting ASC were calculated based on the maximal Elispots on days 1, 2, or 3. Each figure is representative of >3 different experiments
