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目的建立椎间盘髓核细胞(NPCs)和骨髓间充质干细胞(BMSCs)体外聚乳酸-乙醇酸共聚物(PLGA)支架三维共培养系统。方法分别培养乳兔NPCs和BMSCs至原代融合和第3代,将两种细胞按2∶1(BMSCs∶NPCs)比例混匀,以6×106/ml的终浓度分别接种于培养板作为平面共培养组(A组)和PLGA支架(直径10mm,厚3mm)作为三维共培养组(B组);每组各接种6例标本,每例接种细胞悬液100μl,体外培养2周,利用倒置显微镜、扫描电镜进行细胞形态学观察;MTT法测定各组细胞增殖情况。结果细胞与材料支架黏附良好。经过2周的培养,A组和B组细胞均能够增殖;与A组相比,B组细胞增殖率增高(P<0.01)。结论共培养的NPCs和BMSCs均能够与PLGA支架很好地黏附并增殖;PLGA支架三维共培养NPCs和BMSCs较平面共培养更能促进细胞增殖。
OBJECTIVE: To establish a three-dimensional co-culture system of polylactic-co-glycolic acid (PLGA) scaffolds with intervertebral disc nucleus pulposus cells (NPCs) and bone marrow mesenchymal stem cells (BMSCs) Methods Rabbit NPCs and BMSCs were cultured to primary fusion and passage 3, respectively. The two kinds of cells were mixed according to the ratio of 2: 1 (BMSCs: NPCs) and were inoculated into the culture plate at the final concentration of 6 × 106 / ml respectively. Co-culture group (group A) and PLGA scaffold (diameter 10mm, thickness 3mm) as three-dimensional co-culture group (group B); each group were inoculated with 6 specimens, each inoculated with 100μl cell suspension, cultured in vitro for 2 weeks, The morphological changes of the cells were observed by microscope and scanning electron microscope. The proliferation of each group was measured by MTT assay. Results The cells adhered well to the material scaffolds. After 2 weeks of culture, the cells in group A and group B were able to proliferate. Compared with group A, the cell proliferation rate in group B was increased (P <0.01). Conclusions Both co-cultured NPCs and BMSCs can adhere and proliferate well with PLGA scaffolds. Three-dimensional co-culture of NPCs and BMSCs in PLGA scaffolds can promote cell proliferation more effectively than in-plane co-culture.