Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (1): 41-46.doi: 10.3969/j.issn.2095-4344.1524

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Differentiation of human umbilical cord mesenchymal stem cells into neuron-like cells: a comparison of two induction methods

Wang Wu1, Qi Baojun2, Wu Zhongyan1, Cui Yong1   

  1. 1First Department Orthopedics, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China; 2Department of Orthopedics, Kaifeng Central Hospital, Kaifeng 475000, Henan Province, China
  • Revised:2018-09-05 Online:2019-01-08 Published:2018-11-28
  • Contact: Cui Yong, Associate chief physician, Associate professor, Master’s supervisor, First Department Orthopedics, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China
  • About author:Wang Wu, Doctoral candidate, Attending physician, First Department Orthopedics, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China. Qi Baojun, Master, Physician, Department of Orthopedics, Kaifeng Central Hospital, Kaifeng 475000, Henan Province, China. Wang Wu and Qi Baojun contributed equally to this work.
  • Supported by:

    the Natural Science Foundation of Xinjiang Uygur Autonomous Region, No. 2016D01C239 (to WW)

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Abstract:

BACKGROUND: Stem cells are a promising treatment for nerve injury diseases due to its ability of differentiating into neuron-like cells. However, the differentiation efficacy is low, so we have make an advance based on the traditional induction method.
OBJECTIVE: To explore the potentiality of human umbilical cord mesenchymal stem cells differentiating into neuron-like cells induced by two methods and their merits.
METHODS: Human umbilical cord mesenchymal stem cells were isolated and cultured using tissue explant method. The cell morphology was observed under microscope, and the cultured cells were identified by flow cytometry. Human umbilical cord mesenchymal stem cells were differentiated into neuron-like cell under the neurotrophic factor inducing scheme. Two different methods but using the same inducers (2% B27, 20 μg/L basic fibroblast growth factor and 20 μg/L epidermal growth factor) were utilized. Traditional method: B27, basic fibroblast growth factor, epidermal growth factor and penicillin streptomycin combination were added in the DMEM/F12 medium, 3 days later all-trans retinoic acid was added, followed by 10 days of culture. Advanced method: B27, basic fibroblast growth factor, epidermal growth factor, penicillin streptomycin combination and fetal bovine serum were added in the DMEM/F12 medium, 6 days later the cells underwent trypsinization and re-seeded in the confocal capsule, B27 and fetal bovine serum were removed next day, and all-trans retinoic acid was added, cultured for 14 days. Then the differentiated nerve cells were identified by immunofluorescence detection of neurofilament protein and glial fibrillary acidic protein. Meanwhile, the positive rate of the neuromarkers in the cells was compared between two methods.
RESULTS AND CONCLUSION: Primary human umbilical cord mesenchymal stem cells were isolated and cultured successfully, and highly expressed CD29 and CD105 whereas lowly expressed CD34 and CD45, which were the characteristics of the stem cell. The differentiated cells were presented as nerve cells in the morphology under microscope in both inducing methods. The results of the immunofluorescence indicated that the neurofilament protein and the glial fiber acidic protein in the differentiated cells were all expressed positive in both methods. The positive rates of neurofilament protein and glial fiber acidic protein were 18.73% and 18.01% for the traditional method, while 27.48% and 29.24% for the advanced methods, respectively (P < 0.05). In summary, human umbilical cord mesenchymal stem cells can differentiate into the neuron-like cells induced by two methods. Moreover, the advanced method is better than the traditional method in the morphology of the differentiated cells and the positive rate of neurofilament protein and glial fiber acidic protein. 

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: Umbilical Cord, Mesenchymal Stem Cells, Cell Differentiation, Neurons, Tissue Engineering

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