Fig. 4
From: Development of the membrane ceiling method for in vitro spermatogenesis
The effect of advanced DMEM/F12 (AD) as the in vitro basal medium in spermatogenesis. (A) The ratio of tissue area expansion and (B) the GFP-positive rate for 5 weeks in MEMα or AD-based culture medium. There were no significant differences (P > 0.05). (C) Representative images of mCherry-positive cells observed using an inverted microscope (BZ-X700) after 5 weeks cultivated in MEMα or AD-based culture medium. Scale bar = 50 μm. (D) The number of mCherry-positive cells divided by tissue area (mm2). Each dot shows the average cell number per tissue area of frames. Images were taken at 5 weeks using the tiling function of a microscope (BioStation CT and BZ-X700). *P < 0.05. Fifteen (MEMα) and 18 (AD) tissues were used for quantification. (E) In vitro generated sperm after 5 weeks in AD-based culture medium. The images were taken using an upright microscope (BX53). Flagellated cells with normal (upper panel) or abnormal (lower panel) heads were used for ICSI. Scale bar = 20 μm. (F) A picture of obtained offspring by ICSI using in vitro generated sperm.
