体外培养人退变髓核组织细胞的活力和表型变化

目的建立人退变髓核组织细胞体外培养体系,探索最佳培养条件,为髓核退变机制研究和组织工程研究筛选种子细胞。方法采用酶消化法分离人退变髓核组织细胞,进行单层培养,通过CellCountingKit-8评估细胞活性,采用免疫细胞化学染色等方法比较不同代次细胞形态结构及软骨样表型和增殖活性。结果成功进行细胞单层培养和鉴定,P1～P3代细胞活性和Ⅱ型胶原的表达有明显降低（P〈0.05）。结论 P1、P2和P3代细胞表型保持符合软骨样细胞特征,能够为髓核退变体外研究和组织工程研究提供种子细胞。

The Change of Viability and Phenotype of Human Degenetative Nucleus Pulposus Tissue Cells Cultured in vitro

Objective To establish the two-dimensional culture system of the human degenetative nucleus pulposus tissue cells in vitro,screening compatible seed cells for the mechanism study of nucleus pulposus degeneration and tissue engineering. Methods The human degenetative nucleus pulposus tissue cells were isolated with enzyme digestion and grew as monolayer. The morphous,chondrocyte-like phenotype and proliferative viability were compared among different cell passages through Cell Counting Kit-8 and immunocytochemistry of collagen typeⅡ expression. Results The the two-dimensional culture system in vitro was established and identified successfully. The viability and collagen typeⅡ expression of P3 cells dicreased significantly than that of P1 cells. Conclusion P1,P2 and P3 cells cultured in vitro can maintain chondrocyte-like phenotype and proliferative viability,so can be served as the seed cells for the mechanism study of nucleus pulposus degeneration and tissue engineering.