| 摘 要： | 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
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