Effect of conditioned medium from coculture of astrocytes and β-amyloid-induced apoptotic PC12 cells on the differentiation of neural stem cells and the possible mechanism

( views:, downloads: )
YAN Rong(Department of Neurology, Shengjing Hospital Affiliated to China Medical University, Shenyang 110004, China)
LUO Xiao-guang()
BI Guo-rong(Department of Neurology, Shengjing Hospital Affiliated to China Medical University, Shenyang 110004, China)
FENG Juan(Department of Neurology, Shengjing Hospital Affiliated to China Medical University, Shenyang 110004, China)
ZHANG Chao-dong()
Journal Title:
Volume 8, Issue 07, 2009
Key Word:
Astrocytes;Neural stem cells;Neuronal differentiation;Brain-derived neurotrophic factor

Abstract: Objective To observe the effect of the conditioned medium obtained from the coculture of astrocytes and β-amyloid (Aβ1-40)-induced apoptotic PC12 cells on the neuronal differentiation efficiency of neural stem cells (NSCs), and investigate the possible involvement of the neurotrophins proteins including brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF)and neurotrophin-3 (NT-3) in the induced differentiation of the NSCs. Methods PCI2 cells wereinduced by Aβ1-40 for different time lengths (0, 4, 6, 12, and 24 h), and after detection of the apoptotic rates using flow cytometry, the cells were cocultured with astrocytes for 2 days. The astrocyte-conditioned medium (ACM), after analysis of the neurotrophins proteins using erLzyme-linked immunosorbent assay (ELISA), was mixed with DMEM/F12 medium at the proportion of 1:3 to induce the differentiation of the NSCs. The cell differentiation was identified by laser confocal microscopy, NSE immunofluorescent labeling, and neuronal counting. Results Flow cytometry showed that induction by Aβ1-40 for 6 h resulted in the highest apoptosis rate of PC12 cells (P<0.05). BDNF content in the ACM derived from the coculture of astrocytes and the PC12 cells induced for 6 h was significantly increased, and the NSCs induced in this ACM showed the highest neuronal differentiation rates, showing significant difference from those induced by other ACMs (P<0.05). Conclusion The ACM derived from the coculture of Aβ1-40-induced PC12 cells and astrocytes can increase the neuronal differentiation rates of NSCs in vitro, and BDNF in the ACM may play a role in this process.

  • [1]Lovell MA,Geiger H,Van Zant GE,et al.Isolation of neural precursor cells from Alzheimer's disease and aged control postmortem brain[J].Neurobiol Aging,2006,27(7):909-917.
  • [2]Oliveira AA Jr,Hodges HM.Alzheimer's disease and neural transplantation as prospective cell therapy[J].Curr Alzheimer Res,2005,2(1):79-95.
  • [3]Shihabuddin LS,Homer PJ,Ray J,et al.Adult spinal cord stem cells generate neurons after transplantation in the adult dentate gyms[J].J Neurosci,2000,20(23):8727-8735.
  • [4]刘慧,王小军,胡荣,等.星形胶质细胞[J].生理科学进展,2004,35(1):86-91.
  • [5]Perea G,Araque A.Astrocytes potentiate transmitter release at single hippocampal syrmpses[J].Science,2007,317 (5841):1083-1086.
  • [6]Song HJ,Stevens CF,Gage FH.Astroglia induce neurogenesis from adult neural stem cells[J].Nature,2002,417(6884):39-44.
  • [7]Zhou YF,Fang F,Fu JR,et al.An experimental study on astrocytes promoting production of neural stem cells derived from mouse embryonic stem cells[J].Chin Med J,2005,118(23):1994-1999.
  • [8]戴宜武,徐如祥,赵春平,等.神经干细胞发育过程中分化、迁移调控机制研究现状[J].中华神经医学杂志,2003.3(2):140-144.
  • [9]Vicario-Abejon C,Jobe KK,Hazel TG,et al.Function of basic fibroblast growth factor and neurotrophins in the differentiation of hippocampal neurons[J].Neuron,1995,15(1):105-114.
  • [10]Shimazaki T,Arsenijevic Y,Ryan AK,et al.A role for the POU-Ⅲ transcription factor Bin-4 in the regμ Lation of striatal neuron precursor differentiation[J].EMBO J,1999,18(2):444-456.
  • [11]Pencea V,Bingaman KD,Wiegand SJ,et al.Infusion of brain-derived neurotrophic factor into the lateral ventricle of the adult rat leads to new neurons in the parenchyma of the striatum,septum,thulamus,and hypothalamus[J].J Neurosci,2001,21(17):6706-6717.
  • [12]Wang Y,Sheen VL,Maclklis JD.Cortical interneurons upregμ Late neurotrophins in vivo in response to targeted apoptotic degeneration of neighboring pyramidal neurons[J].Exp Neurol,1998,154(2):389-402.
  • [13]Christie SD,Mendez I.Neural transplantation in spinal cord injury[J],Can J Neurol Sci,2001,28(1):6-15.
  • [14]Ahmed S,Reynolds BA,Weiss S.BDNF enhances the differentiation but not the survival of CNS stem cell-derived neuronal precursors[J].J Neurosci,1995,15(8):5765-5778.
WanfangData CO.,Ltd All Rights Reserved
About WanfangData | Contact US
Healthcare Department, Fuxing Road NO.15, Haidian District Beijing, 100038 P.R.China
Tel:+86-010-58882616 Fax:+86-010-58882615 Email:yiyao@wanfangdata.com.cn