| ObjectiveIn vitro, the preliminary mechanism of mouse prostate cancer cells inducing bone marrow mesenchymal stem cells differentiation was discussed. Supplement the mice model and the role of bone marrow mesenchymal stem cells in the development of prostate cancer, so it can provide right direction to further study between certain factor of prostate cancer cells and bone marrow mesenchymal stem cells. For prostate cancer cells by autocrine/fusion mechanism inducing bone marrow mesenchymal stem cells in the direction of prostate cancer cells may provide the theoretical basis.MethodDouble leg femur and fibula was obtained from Anatomic markers with green fluorescent protein in mice, then we got marrow throw washing the cavity in the bone marrow, applied adherent culture screening method and Selected10%fetal bovine serum, appropriate cell inoculation density for cell culture. We indentified mesenchymal stem cells by inversion vary under optical microscope lens cells form, By adipose differentiation reagent culture between bone marrow mesenchymal stem cells using oil red O staining to identify their stem cell properties, Finally by using fluid cytology test CD90, CD105and CD34and CD45index to identify the cell phenotype of bone marrow mesenchymal stem cells. When the cells stick wall more than90%, Separation and amplification will be done. In vitro, non-contact co-culture method was used for MSC and prostate cancer cells in mice. After72h, we observed the morphology of MSC under optical microscope. Through literature, we selected the higher specificity of prostate stem cell antigen (PSCA) indicators as markers for prostate cancer cells in mice, immunohistochemical staining observation was used for the co-culture of MSC in non-contact, so we can have preliminary judgment whether MSC can be induced to the differentiation of prostate cancer cells. ResultThrough attached screening methods for isolation and amplification, we finally got stable represented fibroblasts, long spindle, star and other polytypism cell morphology. After fat cells differentiation, The T3batches of MSC are obviously has a tendency to fat differentiation. Through cell phenotype identification of fluid cytology, the results were CD90positive, positive CD105and CD34negative, CD45negative. In vitro, after72h non-contact culture, the results showed that:1. after co-culture, MSC still had pleomorphic cells form, clear boundary, no obvious abnormality with separate training results.2. The T3batches of MSC undergoing the fat cells differentiation course showed that the oil red O staining is the positive results.3. The result of a high specificity of PSCA index for co-culture of MSC in immunohistochemical staining was a weak positive.ConclusionThrough attached screening method and appropriate cell culture conditions, this experiment successfully got stability batches for cloning prototype sample cell morphological characteristics such as growth and fibroblasts. And by oil red O staining, fluid cytology bone marrow mesenchymal stem cells in mice were verified with stem cells attached growth features.Through in vitro environment non-contact culture prostate cancer cells in mice with MSC, This study preliminary confirmed that after72hours culture, MSC had changed part of the cell phenotype and expressed the PSCA markers. So we can get a preliminary speculation that the mouse prostate cancer cells in co-culture system can secrete a variety of cytokines by paracrine mechanism rather than integration to promote the MSC to prostate epithelial differentiation. Therefore, this study can be speculated that MSC in vitro environment induced by tumor cells factor, can differentiated prostate epithelial cells of tumor. The tumor induced MSCS differentiation theory can help us to understand the role of MSC in tumor formation, for later it can provide a more powerful theory support in many aspects of tumor targeted gene therapy. |
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