Department of Physiology, Christian Medical College, Vellore, Tamil Nadu, India.
Centre for Stem Cell Research, (A unit of InStem, Bengaluru), Christian Medical College, Vellore, Tamil Nadu, India.
Cartilage. 2021 Dec;13(2_suppl):107S-116S. doi: 10.1177/1947603520918635. Epub 2020 May 14.
Articular chondroprogenitors, a suitable contender for cell-based therapy in cartilage repair, routinely employ fetal bovine serum (FBS) for expansion and differentiation. The possibility of transplant rejections or zoonoses transmissions raise a need for xeno-free alternatives. Use of human platelet lysate (hPL), a nutrient supplement abundant in growth factors, has not been reported for human chondroprogenitor expansion thus far. Our aim was to compare the biological profile of chondroprogenitors grown in hPL versus FBS.
Chondroprogenitors were isolated from 3 osteoarthritic knee joints. Following differential fibronectin adhesion assay, passage 0 cells grown in (a) 10% FBS and (b) 10% hPL were considered for assessment of growth kinetics, surface marker expression, gene expression, and trilineage differentiation. Latent transforming growth factor-β1 (TGFβ1) levels were also measured for each culture medium used.
Cellular proliferation was significantly higher in cells grown with hPL ( < 0.01). Surface marker expression was comparable except in CD-146 where hPL group had significantly higher values ( = 0.03). Comparison of mRNA expression revealed notably low values of collagen I, collagen X, aggrecan, and collagen II ( < 0.05). Trilineage differentiation was seen in both groups with higher alizarin red uptake noted in hPL. There were also significantly higher levels of latent TGFβ1 in the medium containing hPL as compared to FBS.
This is the first xeno-free study to affirm that hPL can serve as an optimal growth supplement for expansion of articular chondroprogenitors, although an in-depth assessment of resident growth factors and evaluation of different dilutions of hPL is required to assess suitability for use in translational research.
关节软骨祖细胞是软骨修复细胞治疗的合适候选者,通常使用胎牛血清(FBS)进行扩增和分化。移植排斥或人畜共患病传播的可能性需要无动物来源的替代品。使用富含生长因子的人血小板裂解液(hPL)作为营养补充剂,尚未有报道用于扩增人软骨祖细胞。我们的目的是比较在 hPL 与 FBS 中生长的软骨祖细胞的生物学特性。
从 3 个膝关节骨关节炎关节中分离软骨祖细胞。在差异纤维连接蛋白黏附试验后,对第 0 代细胞进行分析,将在(a)10%FBS 和(b)10%hPL 中生长的细胞用于评估生长动力学、表面标志物表达、基因表达和三系分化。还测量了每种培养基中的潜伏转化生长因子-β1(TGFβ1)水平。
用 hPL 培养的细胞增殖明显更高(<0.01)。表面标志物表达相当,但 CD-146 组 hPL 组的值明显更高(=0.03)。mRNA 表达比较显示,I 型胶原、X 型胶原、聚集蛋白聚糖和 II 型胶原的表达值明显较低(<0.05)。两组均可见三系分化,hPL 组的茜素红摄取量更高。hPL 培养基中潜伏 TGFβ1 的水平也明显高于 FBS。
这是第一项无动物源研究,证实 hPL 可作为扩增关节软骨祖细胞的最佳生长补充剂,尽管需要深入评估内源性生长因子并评估 hPL 的不同稀释度,以评估其在转化研究中的适用性。