caa a22 4500 445823046 CHVBK 20180317145250.0 cr unu---uuuuu 170323e20111201xx s 000 0 eng 10.1007/s10616-011-9379-0 doi (NATIONALLICENCE)springer-10.1007/s10616-011-9379-0 Optimal time for passaging neurospheres based on primary neural stem cell cultures [Elektronische Daten] [Fangling Xiong, Huasong Gao, Yan Zhen, Xue Chen, Weiwei Lin, Jianhong Shen, Yaohua Yan, Xiaodong Wang, Mei Liu, Yilu Gao] Cultured neural stem cells (NSCs) provide a powerful means for investigating central nervous system disease, neuron development, differentiation, and regeneration. To obtain sufficient neurospheres, subculturing is essential following establishment of the primary NSC culture. Passaging the primary neurospheres is a key issue that is often ignored. We evaluated the influence of different passaging schedules on primary cultured NSCs. Passaging was performed on day 5, 7 or 9. We observed more neurospheres with diameters of 200-250μm on day 7 than on day 5 or 9. Prolonging the time of primary culture reduced the cell metabolic activity by the MTT assay and cell proliferation by colony-forming assay and the differentiation to neurons from cells at P2 and later decreased. Additionally, more cells were in G0/G1 phase, and higher expression of p16 INK4a and lower expression of cyclin D1 was found when the time of primary culture was prolonged to 9days compared to 7-days cultures. Thus, in this study, we established that the optimal time for subculturing aggregated NSCs was on day 7 based on the primary culture. Springer Science+Business Media B.V., 2011 Neurosphere nationallicence Neural stem cells nationallicence Primary culture nationallicence Cell passage nationallicence Optimal time nationallicence Xiong Fangling Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut Gao Huasong Department of Neurosurgery, Huashan Hospital affiliated to Fudan University, Shanghai, China aut Zhen Yan Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut Chen Xue Department of Histology and Embryology, Nantong University, Nantong, China aut Lin Weiwei Department of Histology and Embryology, Nantong University, Nantong, China aut Shen Jianhong Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut Yan Yaohua Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut Wang Xiaodong Department of Histology and Embryology, Nantong University, Nantong, China aut Liu Mei Jiangsu Key Laboratory of Neuroregeneration, Nantong University, 19 Qixiu Road, 226001, Nantong, China aut Gao Yilu Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut Cytotechnology Springer Netherlands 63/6(2011-12-01), 621-631 0920-9069 63:6<621 2011 63 10616 https://doi.org/10.1007/s10616-011-9379-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-011-9379-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Xiong Fangling Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut NATIONALLICENCE 700 1- Gao Huasong Department of Neurosurgery, Huashan Hospital affiliated to Fudan University, Shanghai, China aut NATIONALLICENCE 700 1- Zhen Yan Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut NATIONALLICENCE 700 1- Chen Xue Department of Histology and Embryology, Nantong University, Nantong, China aut NATIONALLICENCE 700 1- Lin Weiwei Department of Histology and Embryology, Nantong University, Nantong, China aut NATIONALLICENCE 700 1- Shen Jianhong Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut NATIONALLICENCE 700 1- Yan Yaohua Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut NATIONALLICENCE 700 1- Wang Xiaodong Department of Histology and Embryology, Nantong University, Nantong, China aut NATIONALLICENCE 700 1- Liu Mei Jiangsu Key Laboratory of Neuroregeneration, Nantong University, 19 Qixiu Road, 226001, Nantong, China aut NATIONALLICENCE 700 1- Gao Yilu Department of Neurosurgery, Affiliated Hospital of Nantong University, 20 Xisi Road, 226001, Nantong, China aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/6(2011-12-01), 621-631 0920-9069 63:6<621 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer