FIG. 3.
Analysis of how MSCs affect GBM#1 survival. (A) Expression of growth, angiogenic, and survival factors was analyzed in UCB-MSCs and AT-MSCs by real-time PCR. The expression of factors in UCB-MSCs was normalized to a value of 1 as the standard for each factor. White bar: UCB-MSCs; black bar: AT-MSCs (*P<0.05; **P<0.01). (B) The effect of CXCL12 on GBM#1 was analyzed by FACS. Expression of Annexin V and 7-amino-actinomycin (7-AAD) was determined in the presence or absence of CXCL12. Lower right areas in each (surrounded by bold line) was measured as apoptotic cells according to the manufacturer's instruction. (C) The frequency of 7-AAD/Annexin V+ cells was measured as apoptotic cells by FACS. White bar: GBM#1 alone; hatched-line bar: GBM#1 with recombinant CXCL12; black bar: GBM#1+UCB-MSCs; gray bar: GBM#1+UCB-MSCs with recombinant CXCL12 (**P<0.01). Note that frequency of apoptotic cells after coculture of GBM#1 with UCB-MSCs greatly decreased in the presence of CXCL12. (D) Number of GBM#1 cells after coculture with UCB-MSCs was analyzed with or w/o CXCL12. White bar: GBM#1 alone; hatched-line bar; GBM#1 with recombinant CXCL12; black bar: GBM#1+UCB-MSCs; gray bar: GBM#1+UCB-MSCs with recombinant CXCL12 (**P<0.01). Note that the number of apoptotic cells after coculture of GBM#1 with UCB-MSCs greatly increased in the presence of CXCL12 at the level of GBM#1 alone.