骨关节炎患者髌下脂肪垫前脂肪细胞成脂诱导模型的建立

目的 观察骨关节炎患者髌下脂肪垫前脂肪细胞的原代培养及成脂诱导过程.方法 采用酶消化法分离得到前脂肪细胞,进行细胞形态学观察、CCK-8法测定生长曲线;成脂诱导过程中,油红O染色方法鉴定脂肪细胞、酶联免疫吸附法测定细胞上清脂联素.结果 原代培养的前脂肪细胞呈梭形,与成纤维细胞类似.成脂诱导第3天胞质内出现反光脂质小滴,第21天80%以上的前脂肪细胞可分化为脂肪细胞.细胞上清脂联素随脂肪细胞数量的增多而升高,3周时达到稳定,证实成脂诱导的细胞为功能活跃的脂肪细胞.结论 成功建立了骨关节炎患者髌下脂肪垫前脂肪细胞的原代培养及成脂诱导方法,为研究髌下脂肪垫的内分泌代谢功能及其在骨关节炎发病机制中的作用提供较为理想的细胞模型.

Abstract：

Objective To investigate the primary culture and adipogenic process of pre-adipocytes from infrapatellar fat pad of osteoarthritic patients. Methods The pre-adipocytes were isolated by enzymatic digestion. The morphological changes of cultured cells were observed and the growth curve was drawn by CCK-8 method. During the adipogenic process, the intracytoplasmic lipid of differentiated cells was determined by oil red O staining. And the adiponectin levels in the culture supernatants were measured by ELISA (enzyme-linked immunosorbent assay). Results The primary cultured fibroblast-like cells were spindle-shaped. In the process of adipogenesis, the intracytoplasmic lipid droplets were observed at Day 3and over 80% of the cells differentiated into adipocytes at Day 21. With the increasing number of adipocytes, the adiponectin levels in the culture supemant elevated and peaked at Week 3. The differentiated cells were proven to be adipocytes functioning actively. Conclusion The primary culture and adipogenic process of pre-adipocytes in infrapatellar fat pad of osteoarthritic patients has been successfully established. Thus it may provide an ideal model for the study of endocrine function of infrapatellar fat pad and understanding its role in the pathogenesis of osteoarthritis.

Primary culture and adipogenic process of pre-adipocytes from infrapatellar fat pad of osteoarthritic patients

Objective To investigate the primary culture and adipogenic process of pre-adipocytes from infrapatellar fat pad of osteoarthritic patients. Methods The pre-adipocytes were isolated by enzymatic digestion. The morphological changes of cultured cells were observed and the growth curve was drawn by CCK-8 method. During the adipogenic process, the intracytoplasmic lipid of differentiated cells was determined by oil red O staining. And the adiponectin levels in the culture supernatants were measured by ELISA (enzyme-linked immunosorbent assay). Results The primary cultured fibroblast-like cells were spindle-shaped. In the process of adipogenesis, the intracytoplasmic lipid droplets were observed at Day 3and over 80% of the cells differentiated into adipocytes at Day 21. With the increasing number of adipocytes, the adiponectin levels in the culture supemant elevated and peaked at Week 3. The differentiated cells were proven to be adipocytes functioning actively. Conclusion The primary culture and adipogenic process of pre-adipocytes in infrapatellar fat pad of osteoarthritic patients has been successfully established. Thus it may provide an ideal model for the study of endocrine function of infrapatellar fat pad and understanding its role in the pathogenesis of osteoarthritis.