真菌来源透明质酸对人羊膜间充质细胞的增殖及其向软骨、成骨细胞分化的影响与作用机制

人羊膜间充质细胞(hAMCs)是最具开发潜力的干细胞新资源。透明质酸(HA)有良好的调节增殖分化能力。作为胞外基质(ECM)的主成分，HA用作干细胞增殖分化的调控因子有先天优势。但HA对hAMCs增殖分化的影响及作用机制尚不明确。基于我们的前期研究及文献报道，推测HA是hAMCs增殖及向软骨、成骨细胞分化的潜在调节剂，且Wnt信号在其调控网络体系中扮演了重要角色。为证实这一假说，采用免疫组化、免疫印迹、RT-PCR、RNA干扰、基因芯片等技术，在细胞、基因及蛋白水平上，探讨真菌来源HA对hAMCs的增殖以及分化成软骨、成骨细胞的影响，并揭示Wnt信号通路基因蛋白的变化规律，阐明其作用机制，为完善hAMCs体外增殖分化调控网络体系，及其作为骨及软骨组织工程最佳种子细胞提供实验依据；为hAMCs联合HA用于骨病损细胞治疗、骨及软骨组织工程种植体的构建奠定基础；给真菌来源HA的开发提供了新途径。

Considerable progress has already been made in clinical applications of stem cell research, but still need discovering the regulatory networks of proliferation & differentiation (P&D) in the stem cell development process, a key scientific question for clinical benefit. Human amniotic mesenchyal cells (hAMCs) are recognized as new source for clinical application of stem cells. Hyaluronan (HA) is a good regulator of stem cell P&D. Also, HA, a major component of extracelluar matrix, has natural predominance to regulate the stem cell P&D due to immunogenicity. However, little is known about the influences of HA on the P&D of hAMCs and their acting mechanism. Based on our results and the related research progress on stem cell, we presume that HA can modulate hAMCs proliferation and to differentiated into different cell types such as chondroblast, and osteoblast, in which Wnt signalling plays a vital role in the developmental process. To verify the hypothesis, a series of experimental techniques, including immunohistochemistry, immunoblotting, RT-PCR, RNAi, and Genechip, will be employed to investigate the effects of HA on the P&D of hAMCs and the corresponding changes properties of gene and protein in Wnt pathway, and elucidate their acting mechanisms for the first time. The findings will replenish and enrich the intracellular regulatory networks associating with the P&D of hAMCs, give the further evidences that hAMCs are served as the best seed cells for bone and cartilage tissue engineering. Also, facilitate to repair defects in bone and cartilage, and to construct the implant for bone and cartilage tissue engineering by the combination of hAMCs and HA. At the same time, the findings also provide a new pathway for the exploitation of the fungal-derived HA.