| Part Ⅰ Isolation, Differentiation, and Immunophenotype of Rat Adipose-derived Stem Cells in VitroObjective:To explore the cultivation method and differentiate capacity, identify the immunophenotypes of rat adipose-derived stem cells (ADSCs) in vitro.Methods:ADSCs were cultured by anchorage velocity-dependent separation method after collagenase Ⅰ digestion, then proliferation rates were measured by CCK-8and compared with bone marrow stem cells (BMSCs). The expressions of immunophenotypes were identified by flowcytometrty (FCM), and multi-directional differentiation potentials were identified under specific conditions.Results:The cells with the feature of multiple differentiation potentials can be found in adipose tissues and their proliferation rates were higher than BMSCs significantly. They are characterized by the positive expressions of CD29, CD90, CD106and the negative expressions of CD31, CD34, CD45, CD49, CD73and CD105.Conclusions:ADSCs can differentiate into a variety of phenotypes, which could be easily isolated and cultivated. Their proliferation potentials are superior to BMSCs, and they can long-term proliferate stablely. Part Ⅱ The Effects of Adipose-derived Stem Cells on Erectile Function of Aged Erectile Dysfunction RatsObjective:To explore the effects of adipose-derived stem cells (ADSCs) on the erectile function of aged erectile dysfunction (ED) rats.Methods:Adult rats (5-month old) with normal erectile function and aged rats (25-month old) with erectile dysfunction were selected, and then divided into four groups randomly as adult control group, untreated group, phosphate buffer solution (PBS) treated group and ADSCs treated group. PBS were intracavernous injected to PBS treated group, isovolumetric cell suspensions containing1×106cells were injected to ADSCs treated group, and the other two groups remained untreatment. Intracavernous pressures were monitored2weeks later, and the data were analyzed to evaluate the effects of ADSCs on the erectile function in aged ED rats.Results:1. The ratios of MaxICP/MAP under2.5v,5v,7.5v stimulus in both untreated group and PBS treated group were significantly lower compared with adult control group (P<0.01);2. The ratios of MaxICP/MAP were no significant difference between untreated group and PBS treated group (P>0.05);3. The ratios of MaxICP/MAP in ADSCs treated group were all lower compared with adult control group significantly (P<0.01), but higher than it in untreated group and PBS treated group significantly (P<0.01).Conclusions: ADSCs can significantly improve the erectile function of age-related ED rats. ADSCs would be the promising seed cells for treatment of aged ED patients. Part Ⅲ Adipose-derived Stem Cells restore Erectile Function of Aged Erectile Dysfunction Rats by reversing the structural remodeling of Corpus cavernosumObjective: To investigate whether ADSCs restored erectile function of aged ED rats through reversing structural remodeling of corpus cavernosum.Methods:1. The proliferation rates of ADSCs labeled with CM-DiI were measured by CCK-8, and then the changes of fluorescence intensity were observed within14days and the labeling rates on different time were analyzed.2. Cell suspensions labeled with CM-DiI were intracavernous injected in ADSCs treated group. Two weeks later, corpus cavernosum tissues were incised to identify the expressions of vWF by immunofluorescence and to characterize smooth muscle cells and collagen by Masson staining. The area of vascular endothelial cells (VECs), gross, smooth muscle cells (SMCs) and collagen were quantitative analyzed by Image-Pro Plus6.0software, and the ratios of VECs/gross area and SM/collagen were calculated subsequently for the purpose of assessing the effects of ADSCs on vascular endothelial cells and smooth muscle cells in aged ED rats.Results:1. Fluorescence intensity weakened over time gradually, but, still cleaning visible till14d; the labeling rates in1d,7d and14d were99.22%,97.61%and93.11%, respectively;2. The proliferation rates were no significant difference between labeled and unlabeled cells at any time point (P>0.05);3. Scanty labeled cells were observed in the cavernous tissue2weeks later;4. The expressions of vWF increased in ADSCs treated group, and the sinusoids and Disse space widened, the ratio of VECs/gross area increased and significantly higher than untreated group and PBS group (P<0.01), although significantly lower than adult control group (P<0.01);5. The cavernous smooth muscle cells increased in ADSCs treated group, and the sinusoids and Disse space widened, the ratio of SM/collagen increased and was significantly higher than untreated group and PBS group (P<0.01), although significantly lower than adult control group (P<0.01).Conclusions:Intracavernous administration of ADSCs can effectively regenerate the VECs and SMCs. ADSCs treatment can remodel the corpus cavernosum of aged ED rats. |
PDF Full Text Request