Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (10): 1711-1715.doi: 10.3969/j.issn.1673-8225.2010.10.001

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Differential expression of microRNA-125b in the neuronal differentiation of adipose-derived Flk1+ mesenchymal stem cells

Wang Shi-hua, Bian Chun-jing, Huang Shan, Zhao Chun-hua   

  1. Research Center of Tissue Engineering, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences＆Peking Union Medical College, Beijing  100005, China
  • Online:2010-03-05 Published:2010-03-05
  • Contact: Zhao Chun-hua, Doctor, Doctoral supervisor, Peking Union Medical College, Research Center of Tissue Engineering, Chinese Academy of Medical Sciences, Bejing 100005, China chunhua@public.tpt.tj.cn
  • About author:Wang Shi-hua, Studying for doctorate, Research Center of Tissue Engineering, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences＆Peking Union Medical College, Beijing 100005, China wshawp@ 163.com
  • Supported by:

    Special Foundation of the National Science and Technology Ministry of China (863 Program), No.2006AA02A109*, 2006AA02A115*

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Abstract:

BACKGROUND: The differentiation of mesenchymal stem cells (MSCs) is strictly regulated by both genetic and epigenetic factors. Emerging evidences have demonstrated that microRNA also plays an important role in this process.

OBJECTIVE: To explore the differential expression of microRNA-125b during neuronal differentiation of Flk1+ adipose-derived MSCs (AD-MSCs).

MATERIALS：The fat samples were provided by healthy female volunteers aged 15-35 years and recruited from the Plastic Surgery Hospital affiliated to the Chinese Academy of Medical Sciences.

METHODS: Adult adipose tissues were obtained from healthy volunteers with age of 15-35 years. Using adherence method, Flk1+ MSCs were obtained and the 3rd passage cells were taken in the experiment. Cultured in neuronal induction medium, these MSC were induced to differentiate towards neuronal lineage. The expression of microRNA-125b was examined at days 0, 4, 8 and 12. To explore its role in neuronal differentiation, we need to change its expression. RT-PCR and Taqman real-time PCR were carried out to explore the differentially expression of microRNA-125b during neuronal differentiation of AD-MSCs. The effect of inhibitor on the expression of microRNA-125b was detected.

RESULTS AND CONCLUTION: ①The Flk-1+ MSCs were successfully induced into neuronal differentiation and displayed typical morphological changes 12 days after induction: Most cells retracted their cytoplasm, forming spherical cell body and emitted cellular protrusions. RT-PCR and immunocytochemistry studies confirmed their phenotype with expression of known neuronal cell markers including neurofilament, glial fibrillary acidic protein. ②The expression of microRNA-125b was significant up-regulated during neuronal differentiation. Results of RT-PCR and Taqman real-time PCR were concordance with that of microRNA chip technology. ③Inhibitor could down-regulate microRNA-125b. The results implied that microRNA-125b may play an important role in neuronal differentiation.

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