猫骨髓基质细胞诱导为神经干细胞的实验研究

目的观察猫骨髓基质细胞体外培养及诱导分化情况。方法从猫的骨髓中分离得到骨髓基质细胞(Bone Marrow Stromal cells．BMSC），在体外给予神经干细胞培养基培养，增殖后用分化诱导因子(维甲酸，Retinoic acid，RA)进行诱导分化．倒置相差显微镜下观察活细胞及免疫细胞化学染色情况。结果猫骨髓基质细胞在体外培养中胞体增大，镜下可见丰富的胞浆颗粒，继之出芽，贴壁生长，可形成克隆团，同时可传代培养。这些具有克隆能力的骨髓基质细胞能表达神经干细胞特异性抗原nestin，而且能分化出胶质细胞样细胞和神经元样细胞。结论猫骨髓基质细胞具有干细胞特征，在合适的条件下可扩增、形成克隆并诱导分化出神经胶质细胞和神经元特征细胞，它们可考虑作为神经系统功能缺失细胞移植治疗的种子细胞来源。

The pilot study about the culture and differentiation of the BMSC from the cat

Objective To observe the growth, expansion and differentiation of the cultured bone marrow stromal cells (BMSC) from the cat. Methods The BMSC isolated from the adult cat were cultured with the special culture medium of neural stem cells confected in our lab, then induced by Retinoic acid. The processes of cell culture were observed under inverted phase contrast microscope and identified with immunohistochemical method. Results In the study we found that the BMSC proliferated and generated clones when they were cultured in vitro. Following subculture the cells grew rapidly and differentiated into neuron-like or astrocyte-like cells. We also observed that the cell clones proliferating from stem cells expressed nestin antigen. Conclusion The BMSC from cat have the same characteristics as stem cells do and they can be induced and differentiated into nestin positive cells. If the culture condition was appropriate, they can differentiate into neuron -like or astrocyte-like cells, which may be considered as a kind of perfect seed cells of neural cell therapy.