Defferentiation Interrupted The BMSCs Induced Process And Its Correlation With Tumorigenesis

Recent studies have found that the characteristics of tumor cells growth, metastasis and recurrence are very similar to the basic characteristics of the stem cells. Then some scholars have proposed the cancer stem cell hypothesis, this hypothesis indicated that cancer was cause of these a very small part of tumor cell population which have stem cell properties and unlimited proliferative capacity, what is also called cancer stem cells(Cancer stem cells, CSCs).The hypothesis put forward for us to rethink the origins and nature of the tumor,It also provide a new visual perspective on the future of clinical oncology.While the mature block theory says that the CSCs are produced from stem cells to differentiate normally blocked.On this basis, this experiments using rat bone marrow mesenchymal stem cells(BMSCs) as the research object try to establish the differentiation blocked model of BMSCs into osteoblasts, nerve cells and adipocytes.To investigate the correlation of blocked differentiation of stem cells with tumorigenesis.Specific methods are as follows:1. Choosing the whole bone marrow adherence screening method for isolation of rat BMSCs were cultured in vitro and amplification.2. According to differentiation characteristics of BMSCs, Using different chemical inducer to induce BMSCs to differentiate into osteoblasts, nerve cells, and adipocytes. At different time points were removed the inducing agent to achieve induced differentiation interrupt.Observed all breakpoint cells and differentiated cells morphology and differentiation characteristics.3.The use of soft agar colony formation assay screening the breakpoint cells group which has the ability to form colonies.After the cells were isolated and cultured them to do pathological HE staining, karyotype analysis and experimental animal tumors. To these abnormal tissue which produced by inoculated cells in mice get on histopathological HE staining and immunohistochemical staining.The results are as follows:1.Successfully isolated rat BMSC in vitro and proliferation cultured further to explore the biological characteristics of BMSCs.2.Successfully induced rat BMSCs to differentiate into osteoblasts, nerve cells, and adipocytes, and these cells all exhibited the unique types of mature cells differentiation characteristics.3.The interruption osteogenic induced 4d group, interruption osteogenic induced 5d group, interruption neural induced 44 h group, interruption neural induced 48 h group, interruption adipocytes induced 2d group and interruption adipocytes induced 3d group cells exhibited colony forming ability, After screening these cells to take part in pathology HE staining found cells were occur pathological malignant transformation, By karyotype analysis found that these cells chromosome were abnormalities, animal experiments show that cells which had colony forming ability have different degrees of tumorigenicity.In conclusion, interrupting the normal differentiation of BMSCs would make the cells lose their normal regulation and stop at a certain stage of differentiation to immortalized, subsequently it occured malignant transformation.these cells may be the cancer stem cells.