Chinese Journal of Tissue Engineering Research ›› 2016, Vol. 20 ›› Issue (29): 4328-4333.doi: 10.3969/j.issn.2095-4344.2016.29.010

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Optimization of culture conditions for oligodendrocytes of the rat cerebral cortex

Yang Kai1, 2, 3, Li Yi-peng4, Liu Ying-fu5, Cheng Yuan-chi1, 2, 3, Tang Feng-wu3, Liang Bing3, Xu Zhong-wei4, Chen Xu-yi2, 3   

  1. 1Postgraduate Training Basement, the Affiliated Hospital of Logistics University of Chinese People’s Armed Police Force, Jinzhou Medical University, Jinzhou 121000, Liaoning Province, China; 2Institute of Traumatic Brain Injury and Neurology, the Affiliated Hospital of Logistics University of Chinese People’s Armed Police Force, Tianjin 300162, China; 3the Affiliated Hospital of Logistics University of Chinese People’s Armed Police Force, Tianjin 300162, China; 4Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; 5Logistics University of Chinese People’s Armed Police Force, Tianjin 300162, China
  • Received:2016-04-23 Online:2016-07-08 Published:2016-07-08
  • Contact: Chen Xu-yi, M.D., Master’s supervisor, Institute of Traumatic Brain Injury and Neurology, the Affiliated Hospital of Logistics University of Chinese People’s Armed Police Force, Tianjin 300162, China; the Affiliated Hospital of Logistics University of Chinese People’s Armed Police Force, Tianjin 300162, China
  • About author:Yang Kai, Studying for master’s degree, Postgraduate Training Basement, the Affiliated Hospital of Logistics University of Chinese People’s Armed Police Force, Jinzhou Medical University, Jinzhou 121000, Liaoning Province, China; Institute of Traumatic Brain Injury and Neurology, the Affiliated Hospital of Logistics University of Chinese People’s Armed Police Force, Tianjin 300162, China; the Affiliated Hospital of Logistics University of Chinese People’s Armed Police Force, Tianjin 300162, China
  • Supported by:

    the National Natural Science Foundation of China, No. 11102235; Tianjin Science and Technology Support Project, No. 14ZCZDGX00500; a grant from Health Bureau of Tianjin, China, No. 2013KZ134, 2014KZ135; the Seed Fund of the Affiliated Hospital of Logistics College of Chinese People’s Armed Police Force, No. FYM201417, FYM201432, FYM201542, WHJ2015018; Doctoral Startup Fund of Logistics University of Chinese People’s Armed Police Force, No. WHB201417, WHB201514; Open Fund of Center Laboratory of Logistics University of Chinese People’s Armed Police Force, No. 2015ZXKF01

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Abstract:

BACKGROUND: Oligodendrocytes are mostly differentiated from oligodendrocyte precursor cells. A suitable medium and cell seeding density have a significant impact on the process of the isolation of oligodendrocyte precursor cells to obtain oligodendrocytes.
OBJECTIVE: To explore the optimization of oligodendrocyte culture conditions.
METHODS: Oligodendrocyte precursor cells isolated from the newborn rats 48 hours after birth were cultured in DMEM/high glucose medium or DMEM/F12 medium using seeding densities of 2×104 cells/cm2, 4×104 cells/cm2, 8×104 cells/cm2, 16×104 cells/cm2, 32×104 cells/cm2, and 64×104 cells/cm2, respectively. Oligodendrocyte precursor cells were induced to differentiate into oligodendrocytes at 72 hours after cell adhesion. Morphology of differentiated oligodendrocyte precursor cells were observed under a light microscope, and the differentiation results were identified by immunofluorescence staining after 7-day induced differentiation.
RESULTS AND CONCLUSION: Morphology of oligodendrocyte precursor cells were recognized when cultured in DMEM/high glucose medium or DMEM/F12 medium using seeding densities of 2×104 cells/cm2, 4×104 cells/cm2, and 8×104 cells/cm2, respectively. Immunofluorescence staining showed that myelin basic protein-positive cells were found after 7-day induced differentiation, and the positive cell number were 16.40±3.30, 49.95±2.33, and 76.95±4.86 in DMEM/F12 medium, and 12.65±2.53, 32.10±1.17, and 54.05±1.56 in DMEM/high glucose medium (P < 0.05). These findings indicate that DMEM/F12 medium is more suitable for culturing oligodendrocyte precursor cells compared with DMEM/high glucose medium to some extent. The number of differentiated oligodendrocytes was gradually increased with the enhanced seeding density of oligodendrocyte precursor cells, and the seeding densities from 4×104 to 8×104 cells/cm2 were appropriate for the observation of cell morphology.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Oligodendroglia, Cells, Cultured, Rats

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