Stem Cell Institute, University of Science, VNUHCM, Ho Chi Minh City, Vietnam.
Laboratory of Stem Cell Research and Application, University of Science, VNUHCM, Ho Chi Minh City, Vietnam.
Adv Exp Med Biol. 2019;1084:45-60. doi: 10.1007/5584_2017_133.
In vitro production of tissues or tissue engineering is a promising approach to produce artificial tissues for regenerative medicine. There are at least three important components of tissue engineering, including stem cells, scaffolds and growth factors. This study aimed to produce cartilage tissues in vitro from culture and chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells (BMMSCs), induced by chondrogenesis medium, on biodegradable polycaprolactone (PCL) scaffolds. BMMSCs were isolated from rabbit bone marrow according to the standard protocol. The adherence, proliferation and differentiation of BMMSCs on scaffolds were investigated using two scaffold systems: PCL scaffolds and collagen-coated PCL (PCL/col) scaffolds. The results showed that BMMSCs could attach and grow on both PCL and PCL/col scaffolds. However, the adhesion efficacy of BMMSCs on the PCL/col scaffolds was significantly better than on PCL scaffolds. Under induced conditions, BMMSCs on PLC/col scaffolds showed increased aggrecan accumulation and upregulated expression of chondrogenesis-associated genes (e.g. collagen type II, collagen type I, aggrecan and collagen type X) after 3, 7, 21 and 28 days of induction. These in vitro cartilage tissues could form mature chondrocyte-like cells after they were grafted into rabbits. The results suggest that use of BMMSCs in combination with polycaprolactone scaffolds and chondrogenesis medium can be a way to form in vitro cartilage tissue.
体外组织生产或组织工程是一种有前途的方法,可以生产用于再生医学的人工组织。组织工程至少有三个重要组成部分,包括干细胞、支架和生长因子。本研究旨在通过软骨形成培养基,从兔骨髓来源的间充质干细胞(BMMSCs)的培养和软骨分化,在可生物降解的聚己内酯(PCL)支架上体外生产软骨组织。BMMSCs 是根据标准方案从兔骨髓中分离出来的。使用两种支架系统:PCL 支架和胶原蛋白涂覆的 PCL(PCL/col)支架,研究了 BMMSCs 在支架上的附着、增殖和分化。结果表明,BMMSCs 可以附着和生长在 PCL 和 PCL/col 支架上。然而,BMMSCs 在 PCL/col 支架上的粘附效果明显优于 PCL 支架。在诱导条件下,PCL/col 支架上的 BMMSCs 在诱导 3、7、21 和 28 天后,聚集蛋白聚糖的积累增加,软骨形成相关基因(如 II 型胶原、I 型胶原、聚集蛋白聚糖和 X 型胶原)的表达上调。这些体外软骨组织在移植到兔子体内后可以形成成熟的软骨细胞样细胞。结果表明,使用 BMMSCs 结合聚己内酯支架和软骨形成培养基可以是形成体外软骨组织的一种方法。