脂肪来源干细胞体外成骨和成脂及成神经的诱导分化研究

Adipose-Derived Stem Cells in Vitro and Osteogenic and Adipogenic Induction of Differentiation Into Neurons

目的:探讨脂肪来源干细胞体外成骨和成脂及成神经的诱导分化情况。方法:选取10只SPF级雄性SD大鼠,将其不同部位的脂肪组织取出,分别采用不同方法对其向成骨、成脂及成神经等方向进行诱导分化并对其结果进行鉴定。结果:ADSC表达中,CD29占(99.11±0.13)%,CD44占(95.94±0.71)%,CD45占(0.12±0.09)%。经4周的成骨诱导后,茜素红S染色在细胞团中央发现红色钙化结节存在,碱性磷酸酶染色在细胞的胞质内观察到紫红色颗粒,经7d成脂诱导后,油红"O"染色在细胞质内观察到橙红色脂滴;经过6d的神经干培养基诱导后,通过免疫荧光染色证明诱导的Nestin细胞、神经丝蛋白-200以及GFAP等均出现阳性表达。结论:ADSC具备向脂肪、成骨及神经元等细胞进行多向分化的潜能,具有来源广、易于操作、体外增殖快速等优越性,并且不存在免疫排斥及医学伦理学问题,发展前景广阔。

Objective： To investigate adipose derived stem cells in vitro and osteogenic and adipogenic differentiation into neural induction. Methods： 10 SPF male SD rats to different parts of the fat tissue removed, different methods were used for its osteogenic, adip- ogenic and direction into nerve induced differentiation and the results were identified. Results： ADSC expression, CD29 accounted for （99.11± 0.13）%, CD44 accounted for （95.94± 0.71）%, CD45 share （0.12± 0.09）%. After four weeks ofosteogenic induction, Alizarin red S staining in red cell mass of the central calcified nodules found there, alkaline phosphatase staining was observed in the cytoplasm of cells to purple granules, after 7d adipogenic induction, oil red ＂O＂ staining was observed in the cytoplasm orange-red lipid droplets; thro- ugh 6d after induction of neural stem medium, proved by immunofluorescence staining cells induced Nestin, GFAP and neurofilament protein -200 etc appear positive. Conclusion： ADSC have to fat, bone and neuronal cells such as multiple differentiations potential. Has a wide source, easy to operate, fast proliferation and other advantages, and there is no immune rejection and medical ethics issues and broad development prospects.