粒细胞集落刺激因子对大鼠体内外源性人神经干细胞存活分化的影响

中国儿童保健杂志 ›› 2014, Vol. 22 ›› Issue (2): 149-152.

粒细胞集落刺激因子对大鼠体内外源性人神经干细胞存活分化的影响

崔盈¹, 栾佐², 杨印祥², 王彩英³, 古桂雄³

1 无锡市第四人民医院儿科, 江苏无锡 214062;
2 海军总医院儿科, 北京 100037;
3 苏州大学儿科研究所, 江苏苏州 215003

收稿日期:2013-07-15 发布日期:2014-02-10 出版日期:2014-02-10
通讯作者: 古桂雄, E-mail:szggx000@163.com
作者简介:崔盈(1986-), 江西人, 医学硕士, 研究方向为发育脑保护。
基金资助:
江苏省普通高校研究生科研创新计划项目(CX08B_172Z)

Effect of granulocyte-colony stimulating factor on survival of implanted human neural stem cells and the differentiation into oligodendrocytes.

CUI Ying¹, LUAN Zuo², YANG Yin-xiang², WANG Cai-ying³, GU Gui-xiong³.

(1 Department of Pediatrics, Wuxi No.4 People's Hospital, Wuxi, Jiangsu 214062, China; 2 Department of Pediatrics, Navy General Hospital, Beijing 100037, China; 3 Pediatric Research Institute of Soochow University, Suzhou, Jiangsu 215003, China)

Received:2013-07-15 Online:2014-02-10 Published:2014-02-10
Contact: GU Gui-xiong, E-mail:szggx000@163.com

摘要/Abstract

摘要： 目的观察缺氧缺血脑损伤(hypoxic-ischemic brain damage, HIBD)新生大鼠应用粒细胞集落刺激因子(granulocyte-colony stimulating factor, G-CSF)对移植的人神经干细胞(human neural stem cells, hNSCs)体内存活及分化成少突胶质细胞的影响。方法新生1周的Wistar大鼠HIBD模型后分成两组:NSCs移植组(A组, n=24)和NSCs移植+G-CSF治疗组(B组, n=24), 其中B组建模后1 h即开始皮下注射G-CSF, 每天1次, 移植前后连续5 d注射G-CSF。两组均于建模后第2天经脑室移植hNSCs。移植后3 d用TUNEL法检测植入细胞凋亡情况, 并于移植后1周、2周分别用免疫荧光法检测植入细胞在大鼠脑内的存活及向少突胶质细胞方向分化情况。结果移植后3 d, A组大脑皮层见大量的植入细胞处于凋亡状态, 而B组的植入细胞凋亡明显减少(t=68.36, P<0.01)。移植后1周及2周, 可见植入细胞分布以额叶皮层和海马多见, 纹状体亦有少许植入细胞分布, A组的植入细胞数少于B组(t=14.07和2.5, P<0.05)。而移植后1周及2周, hNSCs几乎不能分化成少突胶质细胞。结论 G-CSF能减少植入细胞的凋亡, 提高植入细胞的存活率, 但对植入细胞向少突胶质细胞方向的分化无影响。

关键词: 缺氧缺血脑损伤, 粒细胞集落刺激因子, 神经干细胞移植, 少突胶质细胞, 新生鼠

Abstract: Objective To examine the effect of granulocyte-colony stimulating factor (G-CSF) on survival of implanted human neural stem cells (hNSCs) and the differentiation into oligodendrocytes in neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods HIBD rats, born within a week, were divided into two groups randomly:the single transplantation group (Group A, n=16) and the G-CSF + transplantation group (Group B, n=16), in which G-CSF[(50 μg/kg·d)]was administered 1 hour after performing operation subcutaneously once a day for five days in a row.In both group animals received implantation of hNSCs into the left cerebral ventricles at the second day after the operation.TUNEL staining was used to observe the apoptosis of the implanted cells in the rat brains 3 days post transplantation.While immunofluorescence was used to detect the survival and oligodendroglial differentiation 1 week and 2 weeks after the transplantation. Results Significantly decreased number of apoptosis cells was observed in group B compared to group A in 3 days after hNSCs transplantation (t=68.36, P<0.01).The grafted cells detected by DAPI survived, and generally distributed among cortex, hippocampus, striatum, and other areas.The statistical analysis revealed that DAPI positive cells were more in rats of group B than those of group A (t=14.07 and 2.5 respectively, P<0.05).Differentiated oligodendrocytes of implanted cells could scarcely be seen in either group. Conclusion The survival of implanted cells is increased and apoptosis is reduced with G-CSF treatment, which has no effect on the differentiation of implanted hNSCs into oligodendrocytes.

Key words: hypoxic-ischemic brain damage, granulocyte-colony stimulating factor, neural stem cells transplantation, oligodendrocyte, neonatal rats

中图分类号:

Q954.5

崔盈, 栾佐, 杨印祥, 王彩英, 古桂雄. 粒细胞集落刺激因子对大鼠体内外源性人神经干细胞存活分化的影响[J]. 中国儿童保健杂志, 2014, 22(2): 149-152.

CUI Ying, LUAN Zuo, YANG Yin-xiang, WANG Cai-ying, GU Gui-xiong.. Effect of granulocyte-colony stimulating factor on survival of implanted human neural stem cells and the differentiation into oligodendrocytes.[J]. journal1, 2014, 22(2): 149-152.

参考文献

[1] Euglund U, Fricker-Gates RA, Lundberg C, et al.Transplantation of human neural progenitor cells into the neonatal rat brain:extensive migration and differentiation with long-distance axonal projections[J].Exp Neurol, 2002, 173(3):1-21.
[2] 屈素清, 栾佐, 尹国才, 等.新生鼠缺氧缺血性脑病后经脑室人神经干细胞的实验研究[J].中华儿科杂志, 2005, 43(8):576-579.
[3] Xiao BG, Lu CZ, Hans L.Cell biology and clinical promise of G-CSF:Immunomodulation and neuroprotection[J].J Cell Mol Med, 2007, 11(6):1272-1290.
[4] 栾佐, 尹国才, 胡晓红, 等.人神经干细胞移植治疗重度新生儿缺氧缺血性脑病1例[J].中华儿科杂志, 2005, 43(8):580-583.
[5] Sato Y, Nakanishi K, Hayakawa M, et al.Reduction of brain injury in neonatal hypoxic-ischemic rats by intracerebroventricular injection of neural stem/progenitor cells together with chondroitinase ABC[J].Reprod Sci, 2008, 15(6):613-620.
[6] Liu H, Jia D, Fu J, et al.Effects of granulocyte colony-stimulating factor on the proliferation and cell-fate specification of neural stem cells[J].Neuroscience, 2009, 164(4):1521-1530.
[7] Yutaka N, Masao K, Takahito K, et al.Granulocyte colony-stimulating factor attenuates neuronal death and promotes functional recovery after spinal cord injury in mice[J].J Neuropathol Exp Neurol, 2007, 66(8):724-731.
[8] Wilkins A, Majed H, Layfield R, et al.Oligodendrocytes promote neuronal survival and axonal length by distinct intracellular mechanisms:a novel role for oligodendrocyte-derived glial cell line-derived neurotrophic factor[J].J Neurosci, 2003, 23(12):4967-4974.
[9] Li QM, Fu YM, Shan ZY, et al.MSCs guide neurite directional extension and promote oligodendrogenesis in NSCs[J].Biochem Biophys Res Commun, 2009, 384(3):372-377.
[10] Rivera FJ, Siebzehnrubl FA, Kandasamy M, et al.Mesenchymal stem cells promote oligodendroglial differentiation in hippocampal slice cultures[J].Cell Physiol Biochem, 2009, 24(3-4):317-324.
[11] Hsieh J, Aimone JB, Kaspar BK, et al.IGF-I instructs multipotent adult neural progenitor cells to become oligodendrocytes[J].J Cell Biol, 2004, 164(1):111-122.
[12] Cecile LM, Delphine B, Christophe K, et al.Directing human neural stem/precursor cells into oligodendrocytes by overexpression of Olig2 transcription factor[J].J Neurosci Res, 2009, 87(15):3438-3446.
[13] Chaubey S, Wolfe JH.Transplantation of CD15-enriched murine neural stem cells increases total engraftment and shifts differentiation toward the Oligodendrocyte Lineage[J].Stem Cells Transl Med, 2013, 2(6):444-454.
[14] Modo M, Stroemer RP, Tang E, et al.Effects of implantation site of stem cell grafts on behavioral recovery from stroke damage[J].Stroke, 2002, 33(9):2270-2278.