ABSTRACT
BACKGROUND: Estrogen exerts a negative regulatory role in adipogenic differentiation of adipose stem cells, but there is no report concerning estrogen effects on adipogenic differentiation of adipose stem cells from the adipose capsule of kidney after long-term cryopreservation. OBJECTIVE: To explore the impact of estrogen on adipogenic differentiation of fresh adipose-derived mesenchymal stem cells from the kidney adipose capsule or those after long-term cryopreservation. METHODS: Passage 3 long-term cryopreserved and fresh adipose-derived mesenchymal stem cells from the kidney adipose capsule were divided into four groups, al of which were induced to adipogenic cells by induced fluid: fresh cells + 10-7 mol/L estrogen, cryopreserved cells + 10-7 mol/L estrogen, fresh cells and cryopreserved cells groups. Oil red O staining and adipogenic quantitative detection were performed at 14 days after induced adipogenic differentiation. RESULTS AND CONCLUSION: There were no differences in the morphology and arrangement between the cryopreserved and fresh cells. Both cryopreserved and fresh cells expressed CD29 and CD 44, but did not express CD31. Intracel ular lipid droplets were observed after adipogenic differentiation by oil red O staining, and the cells were positive for oil red O staining. The adipogenic volume comparison among the four groups was detected on day 14 after adipogenic differentiation, and the absorbance values showed significant difference between the fresh cells and fresh cells + estrogen groups, as wel as between the cryopreserved cells and cryopreserved cells + estrogen groups, but no difference between the fresh and cryopreserved cells groups. It is proved that low-dose estrogen can inhibit the adipogenic differentiation of long-term cryopreserved adipose-derived mesenchymal stem cells from the kidney adipose capsule; however, there is no significant difference between passage 3 long-term cryopreserved and fresh adipose-derived mesenchymal stem cells from the kidney adipose capsule in adipogenic differentiation.
ABSTRACT
Objective To explore the effect of Smad7 gene transfer on tubular cell cycle arrest, tubular apoptosis and fibronectin (FN) synthesis by transforming growth factor ? 1(TGF ? 1) induced. Methods Mouse Smad7 gene was transfected into renal tubular cells in primary cell culture by using Tfx 50 cationic liposome. Tubular cell proliferation was measured by MTT and cell cycle was observed by flow cytometry. The level of FN secretion in the supernatant was determined by ELISA. Results At 48 h after administration of 10 ng/ml TGF ? 1 to renal tubular cells, cell proliferation declined, G 0/G 1 arrested in cell cycle, and cell FN secretion increased significantly. These abnormalities were attenuated by liposome mediated Smad7 gene transfection. Conclusion Transfection and expression of Smad7 can markedly inhibit TGF ? 1 responses in renal tubular cells, which is helpful for further studies of Smad7 gene therapy in vivo .