Sun Tianwei, Yang Zhiming, Xie Huiqi
Division of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, PR China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2005 May;19(5):364-8.
To explore the possibility of constructing tissue engineering muscles by combining allogeneic myoblasts with small intestinal submucosa (SIS) in rabbits.
A large number of purified myoblasts were obtained with multi-procedure digestion and repeated attachment method from skeletal muscles taken from extremities of immature rabbits which were born 7 days ago. The myoblasts were labeled with BrdU, and then combined with SIS to construct tissue engineering muscles. This kind of tissue engineering muscles were grafted into the gastrocnemius muscle defect (1.5 cm in length, 1.0 cm in width) of fifteen rabbits as the experimental group. The SIS was grafted into the same position in the control group. The rabbits were sacrificed 4, 6, 8 weeks after operation. The tissue engineering muscles were evaluated by macroscopic, histological and immunohistochemical observations, and by quantitative analysis of local immunocyte in the grafting site.
Allogeneic myoblasts with SIS were combined perfectly in vitro. The SIS was connected tightly to surrounding skeletal muscles and inflammation response was obvious 4 weeks after grafting. The SIS began to break down and inflammation response became slight 6 and 8 weeks after operation. Compared with that of 8th week, the quantitative analysis of local immunocyte in 4th and 6th week in both experimental and control group has significance (P<0.05). Newly formed muscle tissues were found around SIS in the experimental group in 4th, 6th, and 8th week. Expression of BrdU and myosin immunohistochemical staining were positive in the experimental group and negative in the control group.
Tissue engineering muscles of rabbits which are constructed by combining allogeneic myoblasts with SIS can survive and proliferate.
探讨将同种异体成肌细胞与小肠黏膜下层（SIS）相结合构建兔组织工程肌肉的可能性。
采用多步骤消化和反复贴壁法，从出生7天的未成熟兔四肢骨骼肌中获取大量纯化的成肌细胞。将成肌细胞用5-溴脱氧尿嘧啶核苷（BrdU）标记，然后与SIS相结合构建组织工程肌肉。将这种组织工程肌肉移植到15只兔的腓肠肌缺损处（长1.5 cm，宽1.0 cm）作为实验组。对照组将SIS移植到相同位置。术后4、6、8周处死兔子。通过大体观察、组织学和免疫组织化学观察以及对移植部位局部免疫细胞的定量分析来评估组织工程肌肉。
同种异体成肌细胞与SIS在体外完美结合。SIS与周围骨骼肌紧密相连，移植后4周炎症反应明显。术后6周和8周，SIS开始分解，炎症反应减轻。与第8周相比，实验组和对照组第4周和第6周局部免疫细胞的定量分析有统计学意义（P<0.05）。实验组在第4、6和8周时，在SIS周围发现了新形成的肌肉组织。实验组BrdU和肌球蛋白免疫组织化学染色呈阳性，对照组呈阴性。
将同种异体成肌细胞与SIS相结合构建的兔组织工程肌肉能够存活并增殖。
