CAJ | 학술논문

目的：本研究旨在探讨大鼠肌肉源性干细胞分离培养方法，及向成骨细胞分化的能力，并与传统的骨髓间充质干细胞相比较。方法从大鼠双下肢处获取骨骼肌肉组织，胶原酶消化分离，培养出肌肉源性干细胞。抽取骨髓组织，经过分离，培养出骨髓间充质干细胞。两种干细胞经基础培养基传至第二代时改换诱导培养基，向成骨细胞分化，采用CCK?8法检测两种细胞的增殖能力，矿化结节染色及RT?PCR检测比较向成骨细胞分化能力。结果两种细胞经过诱导后，均可以向成骨细胞方向分化。CCK?8检测显示肌肉源性干细胞细胞活性更强，矿化结节染色也有更多的结节形成。RT?PCR检测显示两种细胞ALP、Osteocalcin和Osteopontin均有表达，但肌肉源性干细胞表达量更高。结论大鼠骨骼肌肉组织可以成功分离培养出肌肉源性干细胞，通过诱导能够较好地向成骨细胞方向分化，有希望成为组织工程理想的种子细胞来源，成为骨髓间充质干细胞一种可供选择的替代。
목적：본연구지재탐토대서기육원성간세포분리배양방법，급향성골세포분화적능력，병여전통적골수간충질간세포상비교。방법종대서쌍하지처획취골격기육조직，효원매소화분리，배양출기육원성간세포。추취골수조직，경과분리，배양출골수간충질간세포。량충간세포경기출배양기전지제이대시개환유도배양기，향성골세포분화，채용CCK?8법검측량충세포적증식능력，광화결절염색급RT?PCR검측비교향성골세포분화능력。결과량충세포경과유도후，균가이향성골세포방향분화。CCK?8검측현시기육원성간세포세포활성경강，광화결절염색야유경다적결절형성。RT?PCR검측현시량충세포ALP、Osteocalcin화Osteopontin균유표체，단기육원성간세포표체량경고。결론대서골격기육조직가이성공분리배양출기육원성간세포，통과유도능구교호지향성골세포방향분화，유희망성위조직공정이상적충자세포래원，성위골수간충질간세포일충가공선택적체대。
Objective To investigate the isolation and culture ,and osteogenic differentiation of rat muscle derived stem cells ,and to compare those with the conventional bone marrow mesenchymal stem cells. Methods The skeletal muscle tissues of rat lower limbs were obtained and isolated by collagenase digestion to culture muscle derived stem cells. The bone marrow tissues were obtained and isolated to culture bone marrow mesenchymal stem cells. After transformed to the second generation,the two kinds of stem cells were cultured from basic medium to induction medium,and were differentiated to osteoblasts. The proliferation of the two stem cells was determined by CCK?8 method. Mineralized nodules staining and RT?PCR test were used to compare the osteogenic differentiation of the two stem cells. Results Both of the muscle derived stem cells and the conventional bone marrow mesenchymal stem cells could be induced to osteogenic differentiation. Higher cell activity and more nodule formation of the muscle derived stem cells were determined by CCK?8 method and the mineralized nodules staining,respectively. The expression of ALP, osteocalcin and osteopontin was found in both of the two stem cells by RT?PCR test,whereas higher expression was found in muscle derived stem cells. Conclusions Muscle derived stem cells can be successfully isolated and cultured from rat skeletal muscle tissue,and well induced to osteogenic differentiation,which might be an ideal source of seed cells for tissue engineering,as an alternative to bone marrow mesenchymal stem cells.