血管外皮细胞的生物学特性及复合细胞外基质仿生支架修复关节软骨损伤

There are 206000000 arthritis patients in China, 1.88 times the population of the three provinces in Northeast China. There need effective intervention to arthritis immediately. The applicant as the head specialist of the "critical technology research and development in tissue engineering", revealing that stem cells cultureed in vitro (forbidden by medical legislation) as tissue engineered seed cells applying in clinic is illegal, and it is the bottleneck of clinical transformation, in process of the "863" topic. It is reported that we can get the pericyte (owed character of stem cell) from adipose tissue easily through lipsuction, and acquire enough to apply in clinic one time (accordance with medical legislation). Is it scientific that use pericyte to repair cartilage defect? Therefore, we carry out practicably fundamental research concerning pericyte derived from adipose tissue as follows: ⑴the extraction and purification of pericyte; ⑵the cell biological characteristics and identification (including the immune characterization) of pericyte in vitro and in vivo;⑶detection of cytokines to evaluate the chondrogenisis of the pericyte; ⑷ the gene test and compare the cell biological information; ⑸the evaluation of the cells in vivo and in vitro chondrogenic potential characteristics; ⑹to evaluate the ability of the cell combined with the cartilage acellular matrix scaffolds to repair injured articular cartilage of weight-bearing area of animal;⑺The biological characteristic changes of cells before and after frozen storage. The aim is to provide the scientific basis for clinical application of pericytes including the seed ceels for tissue engineered cartilage and the other cell therapy.

中国关节炎患者2.06亿人，是东北三省人口总数的1.88倍。关节炎的有效干预迫在眉睫。申请人正在以首席专家牵头的"组织工程关键技术研发""863"课题实施过程中发现：体外培养（医疗管理管理法规不允许）的干细胞作为组织工程临床用种子细胞不合法，成为临床转化的瓶颈。据报道：吸脂术非常容易获得脂肪组织的血管外皮细胞（具有干细胞特性），能一次性获得足够用于临床的数量（医疗管理管理法规允许）。血管外皮细胞用于软骨修复是否具有科学性？为此，开展脂肪组织来源的血管外皮细胞的下列实用性基础研究： ⑴提取与纯化；⑵体内外的细胞生物学特性鉴定（包括免疫特性）；⑶细胞因子水平评价其向软骨细胞诱导潜能；⑷基因检测并比较其细胞生物信息；⑸评价其体内外异位成软骨潜能；⑹ 评价其复合脱细胞软骨基质仿生支架修复动物关节软骨损伤能力；（7）细胞冻存前后的生物学特性变化。为将其用于组织工程或细胞治疗的临床应用提供科学依据。

本课题旨在分离培养脂肪组织来源的血管周围干细胞（血管外皮细胞），验证其生物学特性，并复合组织工程脱细胞细胞外基质支架修复关节软骨损伤。课题组按照课题任务书研究内容完成了下列实用性基础研究：大鼠与人血管周围干细胞分离纯化方法的标准化和细胞标记鉴定；验证了其良好的生物学特性包括增殖与成骨、成脂、成软骨分化能力；全面验证了免疫相关分子表达，证实了其良好的免疫学特性；细胞因子水平比较了与脂肪干细胞表达差异并进行生物信息学分析；高通量基因芯片验证了其基因表达并进行生物信息学分析；体内验证了其调控软骨微环境炎症的能力；构建了血管周围干细胞软骨细胞外基质支架复合物并检测其生物相容性；验证了血管周围干细胞软骨细胞外基质支架复合物调节炎症反应的能力；大鼠及兔体内软骨修复实验，组织学、影像学全面评估了血管周围干细胞细胞支架复合物修复动物关节软骨损伤能力；研究了细胞冻存前后的生物学特性变化。为将其用于组织工程或细胞治疗的临床应用提供科学依据。.本课题发表论文7篇，其中SCI论文5篇，国内核心期刊论文2篇。