Igf-1 Induced Neuronal Stem Cell Differentiation To The Oligodendrocyte Cell Line And Its Transplantation In The Treatment Of Spinal Cord Injury, Experimental Studies

Objectives: To obtain neural stem cells from the hippocampus of newborn wister rats using floating culture method, and to study its characters of proliferation, migration and multipotential capacity of differentiation.Methods: The tissue mass was isolated from the hippocampus of newborn wister rats under the anatomical microscope, and was digested with trypsin. After being washed, cells were seeded into T25 Corning tissue culture flasks with three different densities of 4×10~4cells/ml, 4×10~5 cells /ml and 4×10~6 cells /ml, and incubated at 37℃ in a humidified 5% CO2-95% air atmosphere. The culture medium was composed of DMEM/F12, 20 ng/ml bFGF and B27. At day 3 or 4, half of the medium was echanged. Until day 6, neurospheres were collected, mechanically pipetted into single cells, and then passaged. To examine differentiation of NSCs, cells were seeded onto poly -lysine-coated coverslips in 24-well dishes for about 7-10 days. Mouse anti-MAP2 was used for identification of neurons, rabbit anti-MBP was used for oligodendrocytes and for and rabbit anti-GFAP for astrocytes anti-GFAP.Results: The neurospheres was composed of Nestin positive cells which could tolerate passging. The reunion of the cells could be avoided if the cells were seeded with adensity of 4×l0~4cells/ml. The cells could differentiate into neurons, oligodendrocytes and astrocytes. When the neurospheres were seeded onto poly - lysine-coated coverslips and were cultured with bFGF or IGF-1, these cells would migrate further.Conclusions: 1. The cells isolated from the hippocampus of newborn wister rats and cultured could proliferated, have multipotential capacity of differentiation and were...