Subculture of Germ Cell-Derived Colonies with GATA4-Positive Feeder Cells from Neonatal Pig Testes

Figure 2

Long-term culture using old and fresh feeder cells. Old feeder cells (OFCs), which were attached to the wells in passages 0 and 1 after colony collection on days 10 to 12, were used for subculture in passages 1 and 2 with the collected colonies at passages 0 and 1, respectively ((A), (B), and (C)). Fresh feeder cells (FFCs), isolated from the top of the 40% Percoll layer during density separation of total testicular cells, were used for subculture of colonies of passages 1 to 7. GDCs in passages 2, 4, 8, and 13 are shown in panels (G), (H), (I), and (J), respectively. Alkaline phosphatase-stained colonies in passages 0, 1, and 2 are shown in panels (D), (E), and (F), respectively. Alkaline phosphatease-stained GDCs in passages 2, 4, 8, and 13 are shown in panels (K), (L), (M), and (N), respectively. Scale bars indicate 100 μm in all panels. GDC: germ cell-derived colony.