Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (28): 4555-4561.doi: 10.3969/j.issn.2095-4344.2015.28.023

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Establishment and neural differentiation of spinocerebellar ataxia type 3-induced pluripotent stem cell lines

Luo Min, Hu Dan, Niu Xiao-hua, Song Bing, Ou Zhan-hui, Fan Di, Wang Ding, He Wen-yin, Sun Xiao-fang   

  1. Key Lab for Major Obstetric Diseases of Guangdong Province, Experimental Department of Institute of Gynecology and Obstetrics, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, Guangdong Province, China
  • Online:2015-07-02 Published:2015-07-02
  • Contact: Sun Xiao-fang, Master, Professor, Master’s supervisor, Key Lab for Major Obstetric Diseases of Guangdong Province, Experimental Department of Institute of Gynecology and Obstetrics, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, Guangdong Province, China
  • About author:Luo Min, Master, Key Lab for Major Obstetric Diseases of Guangdong Province, Experimental Department of Institute of Gynecology and Obstetrics, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, Guangdong Province, China
  • Supported by:

     the National Natural Science Foundation of China, No. 31171229, U1132005; the Guangzhou Municipal Science and Information Bureau Project, No. 2011Y1-00038.

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Abstract:

BACKGROUND: Spinocerebellar ataxia type 3 (SCA3) is a typical genetic neurodegenerative disease. To establish patient-specific disease models of genetic background contributes to studying the pathogenesis and exploring therapeutic manners.
OBJECTIVE: To observe the effectiveness of neural differentiation of induced pluripotent stem cell lines induced by SCA3 and the stability of CAG copy number.
METHODS: Skin tissue of SCA3 patient was obtained clinically, and specific skin flbroblasts were isolated and  
cultured. Reprogramming fibroblasts could obtain induced pluripotent stem cells. Patient-specific induced pluripotent stem cells, normal person induced pluripotent stem cells (NHF) and embryonic stem cells (ES-10) were induced to differentiate. Flow cytometry was used to compare the efficiency of differentiation. Western blot assay was utilized to detect ataxin-3 protein expression in neurons. Polymerase chain reaction was applied to measure the CAG repeat number of SCA3/ATXN3 gene.
RESULTS AND CONCLUSION: Induced pluripotent stem cells that had identical genetic background to fibroblasts were successfully obtained, and had similar morphology and multi-directional differentiation potential to human embryonic stem cells. Each cell line could differentiate into neural stem cells. The CAG number did not apparently alter before and after reprogramming as well as induction of neuronal differentiation. The effectiveness of the differentiation of induced pluripotent stem cells derived from SCA3 into neural stem cells was lower than that of normal person-derived induced pluripotent stem cells (NHF) and embryonic stem cells (ES-10). These findings demonstrate that reprogramming can successfully establish human induced pluripotent stem cells, and induced the differentiation of above cells into neural stem cells. In the whole process, CAG number did not obviously alter, which was consistent with body cells of patients. 
中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Spinocerebellar Ataxias, Induced Pluripotent Stem Cells, Cell Differentiation, Neurons

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