Mass Production of Early-Stage Bone-Marrow-Derived Mesenchymal Stem Cells of Rat Using Gelatin-Coated Matrix
Figure 3
The effect of gelatin-coated matrix on the proliferation of BM-MSCs. BM-MSCs were from bone-marrow-derived primary cell populations cultured on dishes coated without or with 1% (wt/v) gelatin for 14 days. Subsequently, BM-MSCs derived from each 2D matrix condition per passage were cultured continuously on 100 mm culture dishes coated without or with 1% (wt/v) gelatin by passage 5. A hemocytometer was used to count the number of negatively trypan blue-stained BM-MSCs obtained at each passage, and the doubling time of BM-MSCs per passage was calculated by , where is time to confluence, is the final cell number, and is the initial cell number. Cell cycle status was investigated in BM-MSCs at passage 5 through FACS analysis. As the results, gelatin-mediated significant increases in cumulative total cell number (a) and total cell number (b) were detected after passages 3 and 2, respectively. A significant decrease in doubling time was found after passage 2 in BM-MSC cultures on a 1% (wt/v) gelatin-coated 2D matrix, compared to cells cultured without gelatin (c). In case of cell cycle status, culture of BM-MSCs on a 1% (wt/v) gelatin-coated 2D matrix increased significantly the yield of BM-MSCs in S/G2/M phase and decreased significantly the yield of BM-MSCs in G0/G1 phase, compared to cell cultured without gelatin (d). All data shown are means ± S.D. of five ((a), (b), and (c)) or three (d) independent experiments. *.
