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目的:探索分离、培养大鼠骨髓间充质干细胞,并诱导其向视网膜神经样细胞分化的实验方案。方法:分离原代SD大鼠BMSCs,培养并鉴定。用含hEGF、bFGF、dbcAMP和IBMX的培养基诱导BMSCs向视网膜神经样细胞分化。进一步用视网膜神经干细胞标签Thy1.1和神经节细胞标签MAP2对分化细胞进行检测。结果:本方法所分离培养的细胞经鉴定表达BMSCs表面标记,加入特定诱导剂后向神经样细胞分化,并表达视网膜神经干细胞特异标记。结论:我们培养的确实是骨髓间充质干细胞,用该诱导剂能够将BMSCs诱导分化为视网膜神经样细胞。这种分离培养和诱导方案对不可逆性视网膜视神经病变具有广泛的临床应用前景。
OBJECTIVE: To explore the experimental protocol for isolation and culture of rat bone marrow mesenchymal stem cells and their differentiation into retinal neuroblastoid cells. Methods: Primary SD rat BMSCs were isolated, cultured and identified. BMSCs were induced to differentiate into retinal neuron-like cells with media containing hEGF, bFGF, dbcAMP and IBMX. Differentiated cells were further probed with retinal neural stem cell marker Thy1.1 and ganglion cell marker MAP2. Results: The cells cultured in this way were identified and expressed on the surface of BMSCs. After adding specific inducer, they differentiated into neuron-like cells and expressed the specific markers of neural stem cells. CONCLUSION: The bone marrow mesenchymal stem cells that we cultured are indeed BMSCs that can be induced to differentiate into retinal neuron-like cells. This method of isolation and culture and induction of irreversible retinal optic neuropathy has a wide range of clinical applications.