Immunoblot changes of dystrophin expression in mice after bone marrow stem cells transplantation / 中国组织工程研究

ABSTRACT

AIM: To investigate the immunoblot changeS of dystrophin expression in mdx mice after bone marrow stem cells transplantation. METHODS: The experiment was conducted in the laboratory of Department of Neurology, the First Affiliated Hospital, Sun Yat-sen University from September to December 2004. ①Twenty-five mdx mice of 7-8 weeks old were selected and randomly divided into transplantation 4, 8, 12 and 16 weeks groups and blank control group with 5 mice in each group. Meanwhile, 20 C57 mice of 4 to 6 weeks old were selected as donor mice, and 5 C57 mice aged 10 weeks served as positive control mice. ②All mice were preconditioned with 7 Gy ?-ray. After radiotherapy, 2?107 marrow stem cells per mouse (about 0.3-0.5 mL) were injected into the vena caudalis of mice; 0.3 mL PBS was injected into the blank control group and positive control group. ③The mice in 4 transplantation groups were killed at each time point, and the gastrocnemius was harvested to prepare dystrophin samples. The amount of dystrophin expression was detected by Western blot with GAPDH as control. RESULTS: All 25 mdx mice and 5 C57 mice were involved in the result analysis. Western Blot analysis of dystrophin after transplantation: No dystrophin was detected in the blank control group; in the 4 weeks after transplantation group, only few dystrophin expressions were detected, and the dystrophin/GAPDH was about 0.095?0.267; in the 12 weeks after transplantation group, dystrophin/GAPDH was about 0.218?0.338; dystrophin expressions were increased with time, at 16 weeks after transplantation, the expressions were much more than those at 8 weeks (dystrophin/GAPDH: 0.393?0.385, 0.173?0.284, t =6.062, P