Huang He, Wang Shukui, Gui Jianchao, Shen Haiqi
Department of Orthopaedic Surgery, Nanjing First Hospital, Nanjing Medical University, 68 Changle Road, Nanjing, 210006, Jiangsu, China.
Central Laboratory, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China.
Cytotechnology. 2016 Oct;68(5):2083-103. doi: 10.1007/s10616-016-9949-2. Epub 2016 Jan 28.
The repair of meniscus in the avascular zone remains a great challenge, largely owing to their limited healing capacity. Stem cells based tissue engineering provides a promising treatment option for damaged meniscus because of their multiple differentiation potential. We hypothesized that meniscus-derived stromal cells (MMSCs) may be present in meniscal tissue, and if their pluripotency and character can be established, they may play a role in meniscal healing. To test our hypothesis, we isolated MMSCs, bone marrow-derived stromal cells (BMSCs) and fibrochondrocytes from rabbits. In order to avoid bone marrow mesenchymal stromal cell contamination, the parameniscal tissues and vascular zone of meniscus were removed. The characters of these three types of cells were identified by evaluating morphology, colony formation, proliferation, immunocytochemistry and multi-differentiation. Moreover, a wound in the center of rabbit meniscus was created and used to analyze the effect of BMSCs and MMSCs on wounded meniscus healing. BMSCs & MMSCs expressed the stem cell markers SSEA-4, Nanog, nucleostemin and STRO-1, while fibrochondrocytes expressed none of these markers. Morphologically, MMSCs displayed smaller cell bodies and larger nuclei than ordinary fibrochondrocytes. Moreover, it was certified that MMSCs and BMSCs were all able to differentiate into adipocytes, osteocytes, and chondrocytes in vitro. However, more cartilage formation was found in wounded meniscus filled with MMSCs than that filled with BMSCs. We showed that rabbit menisci harbor the unique cell population MMSCs that has universal stem cell characteristics and posses a tendency to differentiate into chondrocytes. Future research should investigate the mechanobiology of MMSCs and explore the possibility of using MMSCs to more effectively repair or regenerate injured meniscus.
半月板无血管区的修复仍然是一个巨大的挑战，这主要是由于其愈合能力有限。基于干细胞的组织工程因其多向分化潜能为受损半月板提供了一种有前景的治疗选择。我们推测半月板来源的基质细胞（MMSCs）可能存在于半月板组织中，如果能够确定其多能性和特性，它们可能在半月板愈合中发挥作用。为了验证我们的假设，我们从兔子身上分离出MMSCs、骨髓来源的基质细胞（BMSCs）和成纤维软骨细胞。为避免骨髓间充质基质细胞污染，去除半月板旁组织和半月板的血管区。通过评估形态、集落形成、增殖、免疫细胞化学和多向分化来鉴定这三种细胞的特性。此外，在兔半月板中心制造一个伤口，用于分析BMSCs和MMSCs对受伤半月板愈合的影响。BMSCs和MMSCs表达干细胞标志物SSEA-4、Nanog、核仁素和STRO-1，而成纤维软骨细胞不表达这些标志物。在形态上，MMSCs的细胞体比普通成纤维软骨细胞小，细胞核大。此外，已证实MMSCs和BMSCs在体外均能分化为脂肪细胞、骨细胞和软骨细胞。然而，在充满MMSCs的受伤半月板中发现的软骨形成比充满BMSCs的更多。我们发现兔半月板含有具有通用干细胞特征且有分化为软骨细胞倾向的独特细胞群体MMSCs。未来的研究应调查MMSCs的力学生物学，并探索使用MMSCs更有效地修复或再生受损半月板的可能性。
