Figure 4.

Transplantation of hiSCs into Mouse Kidney and Adrenal Gland

(A) hiSCs implanted directly under the kidney capsule as a fibrin clot.

(B–D) H&E analyses of cells directly implanted under the kidney capsule as a fibrin clot. Blood vessels were observed to develop in explants after three weeks when hiSCs were implanted directly under the kidney capsule as a fibrin clot (C, count in D).

(E–H) Expression of SF1 in controls (E and F) and hiSCs (G and H) implanted directly under the kidney capsule.

(I–L) Expression of CYP11A1 in controls (I and J) and hiSCs (K and L) implanted directly under the kidney capsule.

(M and N) H&E analyses of a mouse adrenal gland transplanted for one week with USCs infected with SF1 24 hr earlier at lower (M) and higher (N) magnification. hiSCs can be observed at the cortex/medulla boundary (arrow in N).

(G–T) Cells transplanted into the adrenal can be visualized by the expression of GFP with immunohistochemistry (O and P), GFP indirect immunofluorescence (Q), or by their expression of CYP17A1 via immunohistochemistry (R–T). Arrows in (O)–(Q) and (S) point to transplanted cells. CYP17A1 is absent in mouse adrenal cortex (R and T). ZG, zona glomerulosa; ZF, zona fasciculata. Scale bars: 50 μm in (B), (C), (E), (G), (I), and (K); 25 μm in (F), (H), (J), (L), (N), (P), (Q), (S), (T); and 100 μm (M), (O), (R).

(U–W) Development of an inducible system aimed at generating unlimited amounts of hiSCs. (U) Schematic of the cumate vector generated to infect USCs. In repressed configuration, CymR repressor strongly binds to the cumate operator site (CuO), downstream of the CMV5 promoter. When cumate is present, CymR is released, which enables transgene expression. (V) USCs were infected with the cumate vector, selected with puromycin, and then treated with cumate or vehicle. Cells underwent similar morphologic changes to those observed in Figure 1G. Scale bars, 50 μm; inset, 20 μm. (W) qRT-PCR analyses of STAR expression after seven days of treatment with increasing concentration of cumate, showing induction dose dependently. Data in (W) are represented as mean ± SEM, n = 3.

See also Figure S5.