组织工程化颌下腺细胞与支架材料复合培养最佳细胞浓度的实验研究

Optimal Cell Density of Tissue Engineered Compound of Submandibular Gland Cells and Galatin Sponge

目的 :研究颌下腺细胞与支架材料体外复合培养中 ,细胞接种浓度对复合培养物的影响。方法 :体外原代培养SD鼠颌下腺细胞 ,并传代至第 2代 ,细胞计数 ,按细胞浓度为 10 ,2 0 ,30 ,4 0 ,5 0 ,6 0 ,70× 10 6/ml接种于 5mm× 5mm× 2mm胶原海绵表面 ,并置于 2 4孔培养板内 ,形成颌下腺细胞 胶原海绵复合物。继续培养 ,于接种细胞后 1,3,5 ,7d取材 ;HE染色、免疫组化、扫描电镜观察及细胞计数检测等 ,评价颌下腺细胞 -胶原海绵复合物形成的最佳细胞接种浓度。结果 :5 0 ,6 0 ,70× 10 6/ml细胞浓度的复合物中颌下腺细胞在支架材料上黏附、伸展及生长良好。 10 ,2 0 ,30 ,4 0× 10 6/ml细胞浓度的复合物中颌下腺细胞在支架材料上增殖分化及生长较差。细胞计数显示接种 7d时 ,各组材料细胞数均只有 10 4/ml数量级 ,5 0 ,6 0 ,70× 10 6/ml接种浓度时 ,细胞粘附数量是 10 ,2 0 ,30 ,4 0× 10 6/ml的 2倍。结论 :颌下腺细胞与胶原海绵复合培养最佳接种细胞浓度为 5 0× 10 6/ml。

Objective: Our aim was to investigate the optimal cell density of tissue engineered compound of submandibular gland cells and galatin sponge Methods:Submandibular gland cells of rats were isolated with pancreation digestin, and then cultured,cells were subcutaneously injected into galatin sponge and the densities of cell suspensions were 10,20,30,40,50,60,70×10 6/ml.We counted the number of cells and observed the histological changes with HE staining,immunohistochemical staining, and scanning electronic microscope after 1,3,5,7 days.Results:With cell densities of 50,60,70 ×10 6/ml cells spread better than those with cell densities of 10,20,30,40×10 6/ml with better attachment ,proliferation, and differentiation. Conclusion:The optimal cell density is 50×10 6/ml ,which can be used in tissue engineered compound of submandibular gland cells and galatin sponge.