脂肪干细胞与可吸收明胶海绵的组织相容性

Histocompatibility of adipose-derived stem cells and absorbable gelatin sponge

背景:在骨组织工程中寻找优良的种子细胞和支架材料非常重要。目前尚未见脂肪干细胞与可吸收明胶海绵在体外进行复合培养的相关报道。目的:诱导脂肪干细胞成骨细胞后,观察其在可吸收明胶海绵上的黏附、增殖情况以及两者的生物相容性,以期为可吸收明胶海绵作为脂肪干细胞的有效载体进行体内移植提供实验基础。方法:体外培养脂肪干细胞,通过免疫组织化学及流式细胞术对其进行鉴定。取第3代脂肪干细胞对其成骨诱导分化,将其接种于多聚赖氨酸处理后的可吸收明胶海绵,进行扫描电镜观察,观察其在明胶海绵上的黏附、生长情况。结果与结论:兔脂肪干细胞48 h内呈圆形、椭圆形贴壁生长,以后逐渐呈长梭形贴壁生长,阳性表达CD29、CD44,阴性表达CD33、CD34。在加入成骨诱导液培养3 d后,脂肪干细胞可诱导为成骨细胞,将诱导后的成骨细胞与可吸收明胶海绵复合培养,24 h可见85%以上已黏附生长,48 h见成骨细胞伸出伪足,7 d见成骨细胞已呈片状黏附生长并分泌大量细胞基质,10 d可见可吸收明胶海绵已开始吸收、降解。提示脂肪干细胞与可吸收明胶海绵具有良好的组织相容性,可吸收明胶海绵可以作为脂肪干细胞的生物载体。

： BACKGROUND：It is particularly important to search favorable scaffolds and seed cells in bone tissue engineering. However, no existing studies have reported the co-culture of adipose-derived stem cells and absorbable gelatin sponge. OBJECTIVE：To induce rabbit adipose-derived stem cells to osteoblasts, observe the adhesion, proliferation and biological characteristics of adipose-derived stem cells on absorbable gelatin sponge, and to provide experimental base for absorbable gelatin sponge as an effective carrier of fat stem cells transplantation in vivo. METHODS：Adipose-derived stem cells were cultured in vitro and identified with immunohistochemistry and flow cytometry. The passage 3 adipose-derived stem cells were induced to differentiate into osteoblasts and then seeded onto absorbable gelatin sponge after treatment with polylysine. The adhesion and proliferation of adipose-derived stem cells on scaffolds were observed under scanning electron microscope. RESULTS AND CONCLUSION：The rabbit adipose-derived stem cells were mainly round and oval shaped at 48 hours, and became long spindle shaped after 48 hours. Adipose-derived stem cells were positive for CD29 and CD44, and negative for CD33 and CD34. Adipose-derived stem cells can be induced to differentiate into osteoblasts after culture in the osteogenesis induced fluid for 72 hours. The induced osteoblasts that were co-cultured with absorbable gelatin sponge had an 85%adherence rate at 24 hours. The cells began to extend pseudopodium at 48 hours, and osteoblasts adhered in clumps and secreted a large amount of cellmatrix at 7 days. Absorbable gelatin sponge began to absorb and degrade. Experimental findings indicate that, adipose-derived stem cells have good histocompatibility with absorbable gelatin sponge, and absorbable gelatin sponge can be used as a biological carrier of adipose-derived stem cells.