Tian Chunxiang, Fan Xuejiao, Chen Xiaohe, Deng Li, Qin Tingwu, Luo Jingcong, Li Xiuqun, Lü Qing
Department of Thyroid & Breast Surgery, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2012 Jun;26(6):749-54.
Extracellular matrix is one of the focus researches of the adipose tissue engineering. To investigate the appropriate method to prepare the porcine skeletal muscle acellular matrix and to evaluate the biocompatibility of the matrix.
The fresh skeletal muscle tissues were harvested from healthy adult porcine and were sliced into 2-3 mm thick sheets, which were treated by hypotonic-detergent method to remove the cells from the tissue. The matrix was then examined by histology, immunohistochemistry, and scanning electron microscopy. The toxic effects of the matrix were tested by MTT. Human adipose-derived stem cells (hADSCs) were isolated from adipose tissue donated by patients with breast cancer, and identified by morphology, flow cytometry, and differentiation ability. Then, hADSCs of passage 3 were seeded into the skeletal muscle acellular matrix, and cultured in the medium. The cellular behavior was assessed by calcein-AM (CA) and propidium iodide (PI) staining at 1st, 3rd, 5th, and 7th days after culturing.
Histology, immunohistochemistry, and scanning electron microscopy showed that the muscle fibers were removed completely with the basement membrane structure; a large number of collagenous matrix presented as regular network, porous-like structure. The cytotoxicity score of the matrix was grade 1, which meant that the matrix had good cytocompatibility. The CA and PI staining showed the seeded hADSCs had the potential of spread and proliferation on the matrix.
Porcine skeletal muscle acellular matrix has good biocompatibility and a potential to be used as an ideal biomaterial scaffold for adipose tissue engineering.
细胞外基质是脂肪组织工程的重点研究内容之一。探讨制备猪骨骼肌脱细胞基质的合适方法，并评估该基质的生物相容性。
从健康成年猪获取新鲜骨骼肌组织，切成2 - 3毫米厚的薄片，采用低渗 - 去污剂法去除组织中的细胞。然后通过组织学、免疫组织化学和扫描电子显微镜对该基质进行检测。采用MTT法检测该基质的毒性作用。从乳腺癌患者捐赠的脂肪组织中分离人脂肪来源干细胞（hADSCs），并通过形态学、流式细胞术和分化能力进行鉴定。然后将第3代hADSCs接种到骨骼肌脱细胞基质中，在培养基中培养。培养后第1、3、5和7天，通过钙黄绿素 - AM（CA）和碘化丙啶（PI）染色评估细胞行为。
组织学、免疫组织化学和扫描电子显微镜显示肌肉纤维完全去除，保留基底膜结构；大量胶原基质呈现规则网络状、多孔样结构。该基质的细胞毒性评分为1级，表明该基质具有良好的细胞相容性。CA和PI染色显示接种的hADSCs在基质上具有铺展和增殖的潜力。
猪骨骼肌脱细胞基质具有良好的生物相容性，有潜力作为脂肪组织工程的理想生物材料支架。
