Figure 5

Histochemical and indirect immunofluorescent analyses of GCM biopsies after ADSC transplantation and CTX injury. Biopsies were collected at one, two, three, four and six weeks after ADSC transplantation (indicated to the left of the panels). (A) Hematoxylin and eosin staining (H & E) staining of ADSC-treated and CTX-injured GCM. (B, C) Indirect immunofluorescent detection of ADSC-derived collagen VI was performed using anti-lamin A/C (green) and anti-α1(VI) collagen (red) antibodies. Images were taken from representative sections at low (B) and high (C) magnification, respectively. Progressive, time-dependent spreading of transplanted ADSC within muscle tissue and donation of type VI collagen into perimysium and endomysium is apparent. (D, E) Indirect immunofluorescence detection of the α1(VI) collagen (green) and type IV collagen (red) co-localization at the basement membrane of the ADSC-treated, CTX-injured GCM. Images were taken from representative sections at low (D) and high (E) magnification, respectively. Nuclei were stained with DAPI (blue). Scale bar, 100 μm (low magnification) and 25 μm (high magnification), respectively. (F, G) Indirect immunofluorescence detection of lamin A/C (green) and type IV collagen (blue) in muscle biopsy after 30 days. Arrow points to donor cells (dc). P, perimysium; e, endomysium. (H) Indirect immunofluorescence detection of Pax7 (green), lamin A/C (red) and type IV collagen (blue) in muscle biopsy after 30 days. (I) Indirect immunofluorescence detection of Pax7 (green), α1(VI) collagen (red) and type IV collagen (blue) in muscle biopsy after 30 days. Arrows point to Pax7-positive (green) satellite cells. Scale bar, 100 μm. ADSC, adipose-derived stem cells; CTX, cardiotoxin; DAPI, 4′,6-diamidino-2-phenyl indol; GCM, gastrocnemius muscle.