Gökçe Alper, Yılmaz Ibrahim, Gökay Nevzat Selim, Can Levent, Gökçe Ciğdem
Nişantaşı University, İstanbul, Turkey.
Rational Drug Use Team, Tekirdağ State Hospital, Tekirdağ, Turkey.
Acta Orthop Traumatol Turc. 2014;48(3):313-9. doi: 10.3944/AOTT.2014.2635.
The aim of this study was to test the hypothesis that insulin, human transferrin, and selenous acid (ITS) preparation have positive effects on chondrocyte proliferation and morphology and investigate the biochemical and histological effects of these additive substances in different cell culture media.
Human cartilage-derived cells (hCDCs) were isolated from the cartilage tissue of a 57-year-old woman diagnosed with gonarthrosis. Tissue samples were cultured in Dulbecco's modified Eagle's medium (DMEM) and RPMI-1640. The cells' chondrogenic activities were observed. After serial passagings, cells were divided into 4 groups at the end of the 6th week. On the 14th day, proliferated cells were examined using an inverted microscope with x4, x10, x20 and x40 magnification and microphotographs were taken. Living cell quantity was determined on the first and 14th days using MTS-ELISA cell proliferation assay.
DMEM (without adding ITS premix solution) and RPMI-1640 containing ITS premix solution provide proliferation of the chondrogenic cells. The proliferation and viability of chondrocytes were revealed in this study in the 3rd group (DMEM solution without additives).
It is suggested that the culture medium ingredients play crucial roles on chondrogenic proliferation in osteochondral tissue cultures.
本研究旨在验证胰岛素、人转铁蛋白和亚硒酸（ITS）制剂对软骨细胞增殖和形态具有积极影响这一假设，并研究这些添加物质在不同细胞培养基中的生化和组织学效应。
从一名57岁被诊断为膝关节骨关节炎的女性软骨组织中分离出人软骨来源细胞（hCDC）。组织样本在杜氏改良 Eagle 培养基（DMEM）和 RPMI-1640 中培养。观察细胞的软骨生成活性。经过连续传代后，在第6周结束时将细胞分为4组。在第14天，使用倒置显微镜以4倍、10倍、20倍和40倍放大倍数检查增殖细胞并拍摄显微照片。在第1天和第14天使用MTS-ELISA细胞增殖测定法测定活细胞数量。
DMEM（不添加ITS预混溶液）和含有ITS预混溶液的RPMI-1640可促进软骨生成细胞的增殖。在本研究中，第3组（无添加剂的DMEM溶液）显示出软骨细胞的增殖和活力。
提示培养基成分在骨软骨组织培养中的软骨生成增殖中起关键作用。
