Fig. 3
From: Development of the membrane ceiling method for in vitro spermatogenesis
Optimization of oxygen-permeable materials for device base. (A) Representative images showing the visibility of seminiferous tubules collected from GARN adult mice in the device using PMP or FEP. Images were taken by an inverted microscope (BZ-X700). Scale bar = 100 μm. (B) The ratio of tissue area expansion and (C) the GFP-positive rate for 5 weeks in the device with PDMS, PMP, or FEP base plate or dish. *P < 0.05, ****P < 0.0001, and ns; no significant. (D) Representative images of He-PAS staining of testis sections after culturing in the device with PDMS, PMP, or FEP base plate or dish. rST round spermatids, eST elongating spermatids. Scale bar = 200 μm (upper panel) and 50 μm (lower panel). (E) Live imaging of in vitro spermatogenesis of the same tubule during cultivation on the PMP bottom plate. The images were taken by BioStation CT. St step of spermatids. d culture days. Scale bar = 50 μm and 10 μm (zoomed in). (F) The number of mCherry-positive cells divided by tissue area. Each dot shows the average cell number per tissue area of frames. Images were taken at 5 weeks in the device with PDMS, PMP, and FEP using the tiling function of a microscope (BioStation CT). **P < 0.01, ****P < 0.0001, and ns; no significant. Seventeen (PDMS), 17 (PMP), 8 (FEP) tissues were used for quantification.
