Durgam Sushmitha S, Stewart Allison A, Pondenis Holly C, Gutierrez-Nibeyro Santiago M, Evans Richard B, Stewart Matthew C
Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL 61802, USA.
Am J Vet Res. 2012 Jan;73(1):153-61. doi: 10.2460/ajvr.73.1.153.
To compare in vitro expansion, explant colonization, and matrix synthesis of equine tendon- and bone marrow-derived cells in response to insulin-like growth factor-I (IGF-I) supplementation.
Cells isolated from 7 young adult horses.
Tendon- and bone marrow-derived progenitor cells were isolated, evaluated for yield, and cultured on autogenous cell-free tendon matrix for 7 days. Samples were analyzed for cell viability and expression of collagen type I, collagen type III, and cartilage oligomeric matrix protein mRNAs. Collagen and glycosaminoglycan syntheses were quantified over a 24-hour period.
Tendon- and bone marrow-derived cells required 17 to 19 days of monolayer culture to reach 2 passages. Mean ± SE number of monolayer cells isolated was higher for tendon-derived cells (7.9 ± 0.9 × 10(6)) than for bone marrow-derived cells (1.2 ± 0.1 × 10(6)). Cell numbers after culture for 7 days on acellular tendon matrix were 1.6- to 2.8-fold higher for tendon-derived cells than for bone marrow-derived cells and 0.8- to 1.7-fold higher for IGF-I supplementation than for untreated cells. New collagen and glycosaminoglycan syntheses were significantly greater in tendon-derived cell groups and in IGF-I-supplemented groups. The mRNA concentrations of collagen type I, collagen type III, and cartilage oligomeric matrix protein were not significantly different between tendon- and bone marrow-derived groups.
In vitro results of this study suggested that tendon-derived cells supplemented with IGF-I may offer a useful resource for cell-based strategies in tendon healing.
比较补充胰岛素样生长因子-I(IGF-I)后马肌腱来源细胞和骨髓来源细胞的体外扩增、外植体定植及基质合成情况。
从7匹年轻成年马分离得到的细胞。
分离肌腱和骨髓来源的祖细胞,评估产量,并在自体无细胞肌腱基质上培养7天。分析样本的细胞活力以及I型胶原、III型胶原和软骨寡聚基质蛋白mRNA的表达。在24小时内对胶原和糖胺聚糖的合成进行定量。
肌腱来源细胞和骨髓来源细胞需要17至19天的单层培养才能达到2代。分离得到的单层细胞的平均±标准误数量,肌腱来源细胞(7.9±0.9×10⁶)高于骨髓来源细胞(1.2±0.1×10⁶)。在无细胞肌腱基质上培养7天后,肌腱来源细胞的数量比骨髓来源细胞高1.6至2.8倍,补充IGF-I的细胞比未处理细胞高0.8至1.7倍。肌腱来源细胞组和补充IGF-I的组中,新的胶原和糖胺聚糖合成显著增加。I型胶原、III型胶原和软骨寡聚基质蛋白的mRNA浓度在肌腱来源组和骨髓来源组之间无显著差异。
本研究的体外结果表明,补充IGF-I的肌腱来源细胞可能为肌腱愈合的细胞治疗策略提供有用的资源。