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目的:分离培养人羊水间充质干细胞(AF-MSCs)及向成骨细胞诱导分化。方法:采用两步培养法分离和培养人AF-MSCs,应用化学诱导法向成骨细胞诱导分化。结果:成功分离、培养和传代人AF-MSCs,部分人AF-MSCs表达Oct-4。两步培养法AF-MSCs集落形成率和Oct-4阳性细胞率均高于一步法。AF-MSCs经成骨诱导后von Kossa法测定可见钙结节呈黑色,碱性磷酸酶活性测定为阳性。结论:人羊水中存在MSCs,两步培养法是一种高效、简便实用而且不干扰常规产前诊断程序的方法。AF-MSCs易诱导分化为成骨细胞,且增殖快,成骨能力强。
OBJECTIVE: To isolate and culture human amniotic fluid-derived mesenchymal stem cells (AF-MSCs) and to differentiate into osteoblasts. Methods: Human AF-MSCs were isolated and cultured by two-step culture method. Osteoblasts were induced by chemical induction. Results: AF-MSCs were successfully isolated, cultured and passaged. Some human AF-MSCs expressed Oct-4. The two-step culture method AF-MSCs colony formation rate and Oct-4 positive cells were higher than the one-step method. After osteogenic induction of AF-MSCs by von Kossa method, calcium nodules were observed to be black, and alkaline phosphatase activity was measured as positive. Conclusion: MSCs are present in human amniotic fluid. Two-step culture method is an efficient, simple and practical method that does not interfere with routine prenatal diagnostic procedures. AF-MSCs are easily differentiated into osteoblasts, proliferate rapidly and have strong osteogenic ability.