Cartilage Tissue Formation Using Redifferentiated Passaged Chondrocytes In Vitro
Cartilage Tissue Formation Using Redifferentiated Passaged Chondrocytes In Vitro
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DOI：
10.1089/ten.tea.2008.0004
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发表时间：
2009-03-01
影响因子：
4.1
通讯作者：
Kandel, Rita A.
中科院分区：
文献类型：
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作者：
Ahmed, Nazish;Gan, Lu;Kandel, Rita A.
当关节软骨受到创伤或退行性疾病(如骨关节炎)的损伤时，修复能力有限，这可能会导致疼痛和生活质量下降。利用组织工程学方法开发的生物表面替代物是一种很有前途的软骨修复方法，它将避免使用目前使用的合成植入物进行全关节置换。体外组织生成的一个基本要求是提供足够数量的细胞，而这些细胞很难从稀疏的细胞软骨组织中获得。此前，我们已经证明体外扩增的去分化软骨细胞(P2)与少量的原代软骨细胞(P0)共培养可以诱导传代(P2)细胞的再分化。在这项研究中，我们表明这种再分化不是暂时的变化。共培养4周后，用流式细胞分离法分离P0、P2细胞，再分化的P2细胞单独培养4周。与P2细胞形成的软骨组织相比，重新分化的DP2细胞形成了更厚的软骨组织。免疫组织化学染色显示，新形成的组织含有II型胶原，每个细胞积累的蛋白多糖比P2细胞形成的组织更多。DP2细胞的II型胶原基因表达高于P2细胞，而I型胶原基因表达低于P2细胞。有趣的是，当DP2细胞与P2细胞共培养时，DP2细胞能够发挥与P0细胞相同的效果。综上所述，在适当的培养条件下，再分化传代软骨细胞的行为与原代软骨细胞相似。这种共培养体系方法可以用来增加通过经典单层细胞扩增获得的分化软骨细胞的数量，为软骨组织工程提供了一种获得足够细胞数量的新途径。
Articular cartilage has limited ability for repair when damaged by trauma or degenerative disease, such as osteoarthritis, which can result in pain and compromised quality of life. Biological surface replacements developed using tissue engineering methods are a promising approach for cartilage repair, which would avoid the need for total joint replacement with the synthetic implants used currently. A basic requirement of in vitro tissue generation is a supply of sufficient number of cells, which are difficult to acquire from sparsely cellular cartilage tissue. Previously, we have shown that coculture of in vitro-expanded dedifferentiated chondrocytes (P2) with small numbers of primary chondrocytes (P0) induces redifferentiation in passaged (P2) cells. In this study we show that this redifferentiation is not a transient change. After 4 weeks of coculture, the P0 and P2 cells were separated by flow-associated cell sorting, and the redifferentiated P2 (dP2) were cultured alone for a further 4 weeks. The redifferentiated dP2 cells formed thicker cartilage tissue compared to the tissue generated by P2 cells. The newly formed tissue contained type II collagen as demonstrated by immunohistochemical staining and accumulated more proteoglycan per cell than the tissue formed by P2 cells. The dP2 cells also exhibited higher type II collagen and lower type I collagen gene expression than the P2 cells. Interestingly, dP2 cells were able to exert the same effect as P0 cells when cocultured with P2 cells. In conclusion, under proper culture conditions, redifferentiated passaged chondrocytes behave similarly to primary chondrocytes. This coculture system approach can be used to increase the number of differentiated chondrocytes that can be obtained by classical monolayer cell expansion and represents a novel way to acquire sufficient cell numbers for cartilage tissue engineering.