Department of Neurosurgery, First Affiliated Hospital of Shantou University Medical College, Shantou 515041, Guangdong Province, China.
Department of Hematology, Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, Guangdong Province, China.
Neural Regen Res. 2012 Dec 25;7(36):2907-14. doi: 10.3969/j.issn.1673-5374.2012.36.008.
The present study co-cultured human embryonic olfactory ensheathing cells, human Schwann cells, human amniotic epithelial cells and human vascular endothelial cells in complete culture medium-containing cerebrospinal fluid. Enzyme linked immunosorbent assay was used to detect nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor secretion in the supernatant of co-cultured cells. Results showed that the number of all cell types reached a peak at 7-10 days, and the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor peaked at 9 days. Levels of secreted nerve growth factor were four-fold higher than brain-derived neurotrophic factor, which was three-fold higher than glial cell line-derived neurotrophic factor. Increasing concentrations of cerebrospinal fluid (10%, 20% and 30%) in the growth medium caused a decrease of neurotrophic factor secretion. Results indicated co-culture of human embryonic olfactory ensheathing cells, human Schwann cells, human amniotic epithelial cells and human vascular endothelial cells improved the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor. The reduction of cerebrospinal fluid extravasation at the transplant site after spinal cord injury is beneficial for the survival and secretion of neurotrophic factors from transplanted cells.
本研究将人胚胎嗅鞘细胞、人雪旺细胞、人羊膜上皮细胞和人血管内皮细胞在含有脑脊液的完全培养基于共培养。酶联免疫吸附试验检测共培养细胞上清液中神经生长因子、脑源性神经营养因子和胶质细胞源性神经营养因子的分泌情况。结果表明，所有细胞类型的数量在 7-10 天达到高峰，神经生长因子、脑源性神经营养因子和胶质细胞源性神经营养因子的表达在第 9 天达到高峰。分泌的神经生长因子水平是脑源性神经营养因子的 4 倍，是胶质细胞源性神经营养因子的 3 倍。生长培养基中脑脊液浓度（10%、20%和 30%）的增加导致神经营养因子分泌减少。结果表明，人胚胎嗅鞘细胞、人雪旺细胞、人羊膜上皮细胞和人血管内皮细胞的共培养提高了神经生长因子、脑源性神经营养因子和胶质细胞源性神经营养因子的表达。脊髓损伤后移植部位脑脊液漏减少有利于移植细胞的存活和神经营养因子的分泌。
