Effects of fetal kidney cells on renal functions in rats with IR ARF.

<p>The difference in (A) BUN, (B) serum creatinine and (C) serum NGAL levels in sham operated, saline treated and fetal kidney cells treated groups at different time points (0, 24, 48, 72 and 96 hours after reperfusion). Values expressed Mean±SEM (n = 6). *p<0.05 vs. sham operated group, #p<0.05 vs. saline treated group.</p

Effects of fetal kidney cells on morphological structure, proliferation and apoptosis of tubular epithelial cells in rats with IR ARF.

<p>(A) Kidney section of sham operated animal showing normal architecture of tubules and glomeruli. (B) Kidney section of saline treated animal showing dilated distal convoluted tubule (solid arrow), swollen and necrotic epithelial cells with nuclear changes in the most proximal convoluted tubule (open arrow) and epithelial or hyaline cast material in lumen (solid arrow head). (C) Kidney section of fetal kidney cells treated animal showing signs of recovery as revealed by mild tubular dilatation (open arrow), desquamation of few proximal convoluted tubules and preservation of the integrity of the cellular structure (solid arrow) (20 X). (D) Jablonski grading score of tubular necrosis in saline and fetal kidney cells treated kidneys after 72 hours of fetal kidney cells therapy. (E-G) Representative immunofluorescence photomicrographs (40X) of PCNA staining of kidney sections of sham operated (E), saline treated (F) and fetal kidney cells treated (G) animals. (H) Quantification of PCNA positive cells per HPF. (I-K) Representative immunofluorescence photomicrographs (40X) of TUNEL staining of kidney sections of sham operated (I), saline treated (J) and fetal kidney cells treated (K) animals. (L) Quantification of apoptotic cells per HPF. Values expressed Mean±SEM. (n = 6), *p<0.05 vs. sham operated group, #p<0.05 vs. saline treated group.</p

<i>In vivo</i> tracking of PKH26 positive cells in IR induced damaged kidney.

<p>Representative immunoflourescence photomicrographs (40X) of fetal kidney cells treated kidney showing (A) CK 19, green (B) Hoechst, blue (C) PKH26 labeled cells, red, located in the interstitial spaces and peri-tubular areas of the kidney (D) Overlay of images of (A), (B) and (C). (E) Overlay of images of (A) and (B). (F) Overlay of images of (B) and (C).</p

Effects of fetal kidney cells therapy on expression of growth factors and pro- and anti-inflammatory cytokines in rat kidney.

<p>(A-I) mRNA expression levels of growth factors viz. bFGF (A), BMP-7 (B), VEGF-A (C) and IGF-1(D), inflammatory cytokines viz. IL-1β (E), TNF-α (F), IFN-γ (G) and IL-6 (H), anti-inflammatory cytokine IL-10 (I). (J) Representative immunoblots showing the expression levels of inflammatory markers viz. NF-kB and ICAM-1 in the kidney tissues of sham operated, saline treated and fetal kidney cells treated groups. (K-L) Bar diagrams showing semi quantitative densitometry of the expression of NFκB and ICAM-1. Comparative gene expression ratio alculated by referring each gene to β-actin as an internal control. Densitometric analysis applied for comparison of relative protein expression and represented in densitometric arbitrary units (a. u.). Values expressed Mean±SEM. *p<0.05 vs. sham operated group. #p<0.05 vs. saline treated group.</p

List of primers used for RQ-PCR analysis.

<p>List of primers used for RQ-PCR analysis.</p

Morphology, karyotype and phenotypic characterization of fetal kidney cells.

<p>(A) Representative photomicrograph (10X) of fetal kidney cells in culture, showing spindle-shape and polygonal morphology. (B) The fetal kidney cells showing normal karyotype at 3rd passage (10X). (C) Flow cytometric analysis of fetal kidney cells showing expression of surface markers CD29, CD44, CD73, CD90, CD105, CD24, CD133, VEGFR2, EpCAM, CD45 and MHC class II (green or red lines, detected with FITC- or PE- conjugated antibodies, respectively) with isotype controls (black lines).</p

Effect of fC-MSC on LV functions: Bar diagrams showing ejection fraction, end systolic volume, end diastolic volume, and left ventricular myo-mass, measured at 1 week after MI (before fC-MSC therapy) and 4 weeks after fC-MSC therapy using gated SPECT analysis.

<p>Values shown are mean ± SEM (n = 6); *P<0.05, *P<0.01, *P<0.001 saline group after cell therapy vs before cell therapy (within the group);^#P<0.05,^#P<0.01, ^#P<0.001 fC-MSC group after cell therapy vs before cell therapy (within the group);^†P<0.05, ^†P<0.01, ^†P<0.001 fC-MSC group (after cell therapy) vs saline group (after cell therapy).</p

fC-MSC inhibit the apoptosis in rats with acute myocardial injury.

<p>(a) Representative immunofluorescence photomicrographs of saline and fC-MSC treated hearts 4 weeks after fC-MSC therapy showing (a-i & b-i) Overlay; (a-ii & b-ii) Tunel positive cells (green dye) counter stain with (a-iii & b-iii) Hoechst dye respectively (B) TUNEL apoptotic index showing significant decrease in apoptotic cells in fC-MSC treated compared to saline treated hearts. Values shown are mean ± SEM (n = 6). **P<0.01 fC-MSC treated vs saline treated hearts. (C) Representative immune-blots showing expression of BAX and BCL2 in saline and fC-MSC treated rats and (D) their relative density. Densitometric analysis was applied for comparison of relative protein expression. Values expressed Mean ± SE (n = 6), *P<0.05, **P<0.01, ***P<0.001: fC-MSC treated vs. saline treated group.</p

Morphology and characterization of fC-MSC (A) Representative photomicrograph (10X, 20 µm) of fC-MSC in culture showing spindle shaped morphology.

<p>(B) Representative flow cytometric dot-plots showing that fC-MSC are (a) CD29+/CD45−; (b) CD44+/CD45−; (c) CD73+/CD31−; (d) CD90+/HLA-DR−; (e) CD105+/HLA-DR−. (C) Representative photomicrographs (10X, 20 µm) showing differentiation of fC-MSC into Osteoblasts (a-i: differentiated cells positive for Alizarin red stain, and a-ii: control cells negative for Alizarin red stain) and Adipocytes (b-i: differentiated cells positive for oil red O stain, and b-ii: control cells negative for oil red O stain).</p

Effect of fC-MSC on gene expression of growth factors in rats with acute myocardial injury.

<p>Bar diagrams showing fold change expression of VEGF, b-FGF, IGF-1, and HGF in saline treated and fC-MSC treated ischemic hearts 4 weeks after fC-MSC administration. Values shown are mean ± SEM of 6 experiments, *P<0.05, **P<0.01, ***P<0.001: fC-MSC vs saline treatment.</p