human umbilical blood-derived mesenchymal stem cells into nerve-like cells*

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1 中国组织工程研究与临床康复第 14 卷第 19 期 出版 Journal of Clinical Rehabilitative Tissue Engineering Research May 7, 2010 Vol.14, No.19 Brain-derived neurotrophic factor, ciliary neurotrophic factor and their combination for in vitro differentiation of human umbilical blood-derived mesenchymal stem cells into nerve-like cells* Huang Wei 1, Miao Zong-ning 1, Chen Lei 2, Zhang Xue-guang 2 1 Department of Critical Care Medicine, Third Wuxi, Wuxi , Jiangsu Province, China; 2 Department of Brain Science, General Hospital of Armed Police Forces, Tianjin , China; 3 Institute of Biotechnology, Soochow University, Jiangsu Province Key Laboratory of Stem Cells, Suzhou , Jiangsu Huang Wei, Master, Associate chief physician, Department of Critical Care Medicine, Third Wuxi, Wuxi , Jiangsu Correspondence to: Miao Zong-ning, Doctor, Associate professor, Department of Critical Care Medicine, Third Wuxi, Wuxi , Jiangsu Supported by: Traditional Chinese Medicine Bureau Planning Program of Jiangsu Province, No. HZ07095* Received: Accepted: ( /WL) Huang W, Miao ZN, Chen L, Zhang XG. Brain-derived neurotrophic factor, ciliary neurotrophic factor and their combination for in vitro differentiation of human umbilical blood-derived mesenchymal stem cells into nerve-like cells. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2010;14(19): [ Abstract BACKGROUND: Umbilical cord blood mesenchymal stem cells (UCBMSCs) have been shown to differentiate into nerve-like cells under the condition of in vitro culture. Brain-derived neurotrophic factors (BDNF) combined with ciliary neurotrophic factors (CNTF) at certain concentration can in vitro induce the differentiation of UCBMSCs into high proportion of neuronal-like cells. OBJECTIVE: To investigate the feasibility of BDNF, CNTF and their combination for in vitro differentiation of UCBMSCs into nerve-like cells. METHODS: UCBMSCs of passage 5 were induced to differentiate into nerve-like cells using 5, 10, 20 μg/l BDNF, 5, 10, 20 μg/l CNTF or their combination. A blank control group was set, with no any interventions. Cell morphology was observed under inverted phase contrast microscope. At 1, 3, and 6 days of experiment, immunocytochemical staining for neuron specific enolase and glial fibrillary acidic protein was performed. The proportion of differentiated neuronal-like cells and glial cell-like cells was calculated. RESULTS AND CONCLUSION: After induced differentiation into nerve-like cells, human UCBMSCs exhibited changeable morphology with contracted cell body, enhanced refraction of nucleus, and dendrite- and axon-like structure. Compared with blank control group, BDNF and CNTF could significantly enhance the differentiation proportion of UCBMSCs into nerve-like cells. 20 μg/l BDNF combined with 20 μg/l CNTF yielded highest differentiation proportion of human UCBMSCs into nerve-like cells. These findings indicate that human UCBMSCs can differentiate into nerve-like cells after in vitro induction of BDNF, CNTF or their combination. INTRODUCTION Mesenchymal stem cells (MSCs) have been considered as a good source of stem cells [1-5] due to tremendous self-renewal and multi-lineage differentiation potential. Some curative effects have been made in animal experiments using MSCs to treat ischemic brain injury [6]. Buzanska et al [7-8] reported that umbilical cord blood-derived MSCs (UCBMSCs) can be induced to express the marker of neurons and glial cells. But the methods used to regulate the in vitro differentiation of MSCs into high proportion of nerve-like cells have been rarely reported. This study observed the induced differentiation of human UCBMSCs into nerve-like cells using different concentrations of brain derived neurotrophic factor (BDNF) combined with ciliary neurotrophic factor (CNTF) and further investigated the methods and mechanisms underlying the induced differentiation of UCBMSCs, hopefully providing experimental evidence and theoretical acknowledge for repair of central nervous system injury. MATERIALS AND METHODS Design Controlled observation. Time and setting This study was performed at the Laboratory of Stem Cells, Third Wuxi between December 2007 and December Materials Reagent/instrument DMEM/F12, FITC-labeled goat anti-mouse IgG, TRITC-labeled goat anti-rabbit IgG BDNF, CNTF Paraformaldehyde SABC kit Mouse anti-human neuron specific enolase (NSE), mouse anti-human glial fibrillary acidic protein (GFAP) Inverted phase contrast microscope, fluorescence microscope, digital image acquisition system Source Gibco, USA Sigma, USA Shanghai Chemical Reagent Factory, China Wuhan Boster, China Chemicon, USA Olympus, Japan Methods In vitro culture of human UCBMSCs Harvest, primary culture, passage, and identification of human UCBMSCs were performed according to previously reported methods [9]. Material preparation Following at least five generations of culture, well growing human UCBMSCs were digested with 0.25% 3606

2 Huang W, et al. Brain-derived neurotrophic factor, ciliary neurotrophic factor and their combination for in vitro differentiation of human... trypsin-0.01% EDTA, washed twice with phosphate buffered saline (PBS), centrifuged, collected using DMEM/F12, and then inoculated into a 6-well plate at /L for 4 days for later use. Induced differentiation using growth factors Following culture of DMEM/F12 containing 5% fetal bovine serum, adherent cells were inoculated into 6-well plates at /L. Different concentrations of BDNF, CNTF or their combination were added as follows: blank control group: none added; BDNF group: 5, 10, 20 μg/l BDNF added; CNTF group: 5, 10, 20 μg/l BDNF added; BDNF+CNTF group: 5 μg/l BDNF+ 5 μg/l CNTF, 10 μg/l BDNF+ 10 μg/l CNTF, and 20 μg/l BDNF+ 20 μg/l CNTF added. After culture, cells were identified at 1, 3, and 6 days of experiment. Observation under inverted phase contrast microscope was performed and results were recorded. Immunocytochemistry identification Induced and blank control cells were fixed with 40 g/l paraformaldehyde for 30 minutes, washed twice with PBS, treated with 0.1% Triton X-100 for 20 minutes, and blocked with serum working fluid for 20 minutes at room temperature. Following serum removal and mouse anti-human NSE (1: 1 000) and GFAP (1: 500) antibody addition, cells were treated in the moist box overnight at 4, followed by three PBS washes. Immunocytochemistry was performed as aforementioned. Following addition of goat anti-mouse Ig-FITC and Ig-TRITC antibody, cells were incubated at 37 for 2 hours, washed with PBS three times, observed and photographed through the use of fluorescence microscope. Design, enforcement and evaluation This study was designed by the first author, performed by all authors, and evaluated by the third author. All authors received professional training. Blind method was not employed. the aggregation of NSE/GFAP-positive neurons with brown yellow cytoplasm. Immunocytofluorescent staining revealed NSE/GFAP-positive neurons with green cytoplasm (Figures 1-4). Figure 1 Neuron specific enolase immunohistochemistry of differentiated human umbilical cord blood mesenchymal stem cells ( 100) Figure 2 Neuron specific enolase immunocytofluorescent Main outcome measures Cell growth in all groups; NSE and GFAP-positive cell expression in all groups after induction by different concentrations of BDNF, CNTF or their combination. Statistical analysis All data were statistically processed by the first author using SPSS 10.0 software and expressed as Mean ± SD. A level of P < 0.05 was considered statistically significant. Figure 3 Glial fibrillary acidic protein immunohistochemical RESULTS Induction of different concentrations of BDNF, CNTF or their combination At 2 hours after induction by different concentrations of BDNF, CNTF or their combination, obvious cell morphology change was observed under the optical microscope: cytoplasm of UCBMSCs contracted towards nucleus, presenting with typical appearance of perikaryon. After 3-5 hours, most cells showed neuronal-like cell morphology with round cell body, longer processes, branches at the end of processes, and some adjacent cells interweaved, but cell number did not increase. Three days later, most cells showed bipolar or multipolar neuronal-like morphology, with processes similar to axons or dendrites, some processes interweaved. Immunohistochemical staining revealed Figure 4 Glial fibrillary acidic protein immunocytofluorescent ISSN CN /R CODEN: ZLKHAH 3607

3 Huang W, et al. Brain-derived neurotrophic factor, ciliary neurotrophic factor and their combination for in vitro differentiation of human... NSE-positive expression at different time points following induction by different concentrations of BDNF, CNTF or their combination NSE-positive expression was detected in the BDNF, CNTF or BDNF+ CNTF groups but not in the blank control group. BDNF induced higher proportion of NSE-positive cells at 10 μg/l, 20 μg/l than at 5 μg/l at the same time period (P < 0.05). NSE-positive expression was gradually increased at 1-3 days, and it tended to be stable at 3-6 days. There was no significant difference in NSE-positive expression between 10 μg/l and 20 μg/l BDNF groups (P > 0.05). CNTF produced similar effects on induction of UCBMSCs to BDNF. The combination of BDNF and CNTF at different concentrations yielded significantly higher proportion of NSE positive cells than single use (P < 0.05), and showed gradually increased NSE expression within 1-3 days and stable expression within 3-6 days. The effects of BDNF combined with CNTF were not equal to the effect superposition of each single factor (Table 1). Table 1 NSE-positive cell expression at different time points following induction by different concentrations of BDNF, CNTF or their combination (x _ ±s, %) GFAP-positive cell expression at different time points following induction by different concentrations of BDNF, CNTF or their combination (Table 2) 3608 Group Day 1 Day 3 Day 6 Blank control BDNF 5 μg/l 3.4± ±2.5 a 3.8±2.1 a 10 μg/l 4.9± ±3.3 a 10.9±1.7 a 20 μg/l 5.1± ±2.3 a 12.1±2.1 a CNTF 5 μg/l 3.5± ±2.2 a 4.2±2.8 a 10 μg/l 6.3± ±4.6 a 12.6±2.7 a 20 μg/l 6.8± ±1.8 a 13.3±2.5 a BDNF+CNTF 5 μg/l BDNF+5 μg/l CNTF 23.2± ±1.9 ab 47.6±3.2 ab 10 μg/l BDNF+10 μg/l CNTF 35.3± ±2.7 ab 67.1±2.8 ab 20 μg/l BDNF+20 μg/l CNTF 36.1± ±3.2 ab 68.2±4.2 a NSE: neurone specific enolase; BDNF: brain derived neurotrophic factor; CNTF: ciliary neurotrophic factor. a P < 0.05, vs. day 1; b P < 0.05, vs. BDNF or CNTF group Table 2 GFAP-positive cell expression at different time points following induction by different concentrations of BDNF, CNTF or their combination (x _ ±s, %) Group Day 1 Day 3 Day 6 Blank control BDNF 5 μg/l 1.6± ±2.3 a 3.3±2.0 a 10 μg/l 2.6± ±3.1 a 8.1±1.9 a 20 μg/l 3.3± ±3.3 a 8.3±2.3 a CNTF 5 μg/l 1.1± ±3.6 a 4.2±2.1 a 10 μg/l 4.7± ±2.3 a 9.1±2.6 a 20 μg/l 5.2± ±2.1 a 5.4±2.6 a BDNF+ CNTF 5 μg/l BDNF+5 μg/l CNTF 8.7± ±2.7 ab 20.0±2.4 ab 10 μg/l BDNF+10 μg/l CNTF 18.5± ±3.3 ab 27.1±3.3 ab 20 μg/l BDNF+20 μg/l CNTF 22.1± ±2.6 ab 27.4±2.8 ab GFAP: glial fibrillary acidic protein; BDNF: brain derived neurotrophic factor; CNTF: ciliary neurotrophic factor. a P < 0.05, vs. day 1; b P < 0.05, vs. BDNF or CNTF group GFAP-positive expression was detected in the BDNF, CNTF or BDNF+ CNTF groups but not in the blank control group. BDNF at 10 μg/l, 20 μg/l induced higher proportion of GFAP-positive cells than at 5 μg/l within 3-6 days (P < 0.05); there was no significant difference in GFAP-positive expression between 10 μg/l and 20 μg/l BDNF groups at the same time period (P > 0.05). CNTF produced similar effects on induction of UCBMSCs to BDNF, i.e., at each time point, CNTF or CNTF at 10 μg/l and 20 μg/l induced higher proportion of GFAP-positive cells than at 5 μg/l, and there was no significant difference in GFAP-positive cells between l0 μg/l and 20 μg/l groups (P > 0.05). DISCUSSION This study observed the effects and efficiency of BDNF, CNTF or their combination in induced differentiation of UCBMSCs into nerve-like cells. Immunohistochemistry of NSE and GFAP molecular markers [10] revealed that under the condition of in vitro culture, UCBMSCs could differentiate into nerve-like cells [11] and confirmed the feasibility of acquiring high proportion of neuronal-like cells in vitro. Results from this study demonstrated that BDNF or CNTF could induce UCBMSCs to differentiate into nerve-like cells [12] and produced similar effects in promoting cell differentiation; BDNF combined with CNTF could acquire high proportion of nerve-like cells, and more than 80% of cells expressed NSE and GFAP markers. After 3 days of induction by BDNF, CNTF or their combination, nerve-like cells formed, as confirmed by NSE (a specific marker for neuron) and GFAP (a specific marker for glial cell) expression. Such cells appeared in the sporadic manner initially and in a gradual increasing tendency thereafter. These findings indicate that this protocol is feasible and efficient, and that to enhance the differentiation proportion of nerve-like cells, some improvements should be made in induction regulation in vitro. Nerve growth factors in the culture of cells and tissues have been confirmed to be able to promote neuronal survival and differentiation and regulate axonal and dendrite growth [13]. Some scholars used epidermal growth factors, retinoic acid, and BDNF to induce bone marrow-derived MSCs and found that MSCs could differentiate into neuronal-like cells and differentiated cells expressed nestin, as confirmed by immunohistochemistry. Based on this, this study used BDNF combined with CNTF to induce UCBMSCs and acquired high proportion of neuronal-like cells and immunohistochemistry confirmed that the differentiated cells showed nerve-like morphology and NSE and GFAP expression. Quirici et al [15] reported that there was low-affinity nerve growth factor receptor on the surface of MSCs. All growth factor receptors have tyrosinase activity. Once growth factors bind to correspondent receptors, a series of protein kinases would cause the phosphorylation of many protein substrates, and lead to immediate early gene transcription, which causes gene duplication, transcription and finally cell proliferation and differentiation [16]. BDNF is a basic protein harvested from brain tissue extract, with a relative molecular mass of It is primarily distributed in the central nervous system, such as cerebral cortex, medulla, cerebellum, and hippocampus, and in the peripheral heart, lung, skeletal muscle, and sciatic nerve [17-18]. It provides nutrition to neurons in vivo and in vitro, maintains

4 Huang W, et al. Brain-derived neurotrophic factor, ciliary neurotrophic factor and their combination for in vitro differentiation of human... neuronal survival, promotes axonal elongation, and plays an important role in regulating synapse functions in the hippocampus and visual cortex. BDNF has been considered feasible in treating neurodegenerative diseases due to its role in nervous system development, adult nerve plasticity, and nerve structure integrity [19-20]. BDNF binding to Trk extracellular region activates tyrosine kinase, produces two important second messengers IP3 and DG, which induce Ca 2+ release and activate protein kinase C, respectively [21-23]. CNTF is initially extracted from chicken ciliary body and named by its role in maintaining the survival of chicken parasympathetic ganglions [24-25]. CNTF possesses multiple functions and can promote the survival of various nerve cells. It is the first neurotrophic factor found to maintain the survival of spinal cord motor neurons and the growth of processes in vivo and in vitro and plays an extremely important role in nervous system development, differentiation and nerve injury repair [26-27]. CNTF also can induce neuronal differentiation, inhibit sympathetoblast proliferation and promote its differentiation [28]. Following central nerve system injury, rapidly synthesized CNTF is released into extracellular space and provides nutrition to neurons and glial cells in the injured region, so CNTF is considered a repair factor for central nervous system injury [29-30]. Results from this study also suggest that BDNF combined with CNTF can produce a synergic effect on differentiation of UCBMSCs and yields a higher proportion of neuronal-like cells than BDNF or CNTF induction. To conclude, under the condition of in vitro culture, UCBMSCs can differentiate into nerve-like cells and high proportion of neuronal-like cells can be acquired. REFERENCES [1] Iguchi A, Kobayashi R, Sato TZ, et al. Successful report of reduced-intensity stem cell transplantation from unrelated umbilical cord blood in a girl with chronic active Epstein-Barr virus infection. J Pediatr Hematol Oncol. 2006;28(4): [2] Park KS, Lee YS, Kang KS. In vitro neuronal and osteogenic differentiation of mesenchymal stem cells from human umbilical cord blood. J Vet Sci. 2006;7(4): [3] Kang XQ, Zang WJ, Bao LJ, et al. Differentiating characterization of human umbilical cord blood-derived mesenchymal stem cells in vitro. Cell Biol Int. 2006;30(7): [4] Zhao MZ, Nonoguchi N, Ikeda N, et al. Novel therapeutic strategy for stroke in rats by bone marrow stromal cells and ex vivo HGF gene transfer with HSV-1 vector. J Cereb Blood Flow Metab. 2006;26(9): [5] Gargett CE, Chan RW, Schwab KE. Endometrial stem cells. Curr Opin Obstet Gynecol. 2007;19(4): [6] Yan XH, Chen X. Intravenous transplantation of umbilical blood mesenchymal stem cells after hypoxic-ischemic brain damage in neonatal rat. Zhong guo Xinsheng Erke Zazhi. 2006;21(3): [7] Sanchez-Ramos JR, Song S, Kamath SG, et al. Expression of neural markers in human umbilical cord blood. Exp Neurol. 2001;171(1): [8] Buzańska L, Machaj EK, Zabłocka B, et al. Human cord blood-derived cells attain neuronal and glial features in vitro. J Cell Sci. 2002;115(Pt 10): [9] Huang W, Zhu JZ, Miao ZN, et al. In vitro culture and influencing factors of mesenchymal stem cells derived from human umbilical blood. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2007;11(3): [10] Chan RW, Gargett CE. Identification of label-retaining cells in mouse endometrium. Stem Cells. 2006;24(6): [11] Cervelló I, Simón C. Somatic stem cells in the endometrium. Reprod Sci. 2009;16(2): [12] Yokoyama M, Miwa H, Maeda S, et al. Influence of fetal calf serum on differentiation of mesenchymal stem cells to chondrocytes during expansion. J Biosci Bioeng. 2008;106(1): [13] Sasson IE, Taylor HS. Stem cells and the pathogenesis of endometriosis. Ann N Y Acad Sci. 2008;1127: [14] Wang Y, Crisostomo PR, Wang M, et al. Nitric oxide suppresses the secretion of vascular endothelial growth factor and hepatocyte growth factor from human mesenchymal stem cells. Shock. 2008;30(5): [15] Quirici N, Soligo D, Bossolasco P, et al. Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies. Exp Hematol. 2002;30(7): [16] Lee NK, Sowa H, Hinoi E, et al. Endocrine regulation of energy metabolism by the skeleton. Cell. 2007;130(3): [17] Ginn SL, Fleming J, Rowe PB, et al. Promoter interference mediated by the U3 region in early-generation HIV-1-derived lentivirus vectors can influence detection of transgene expression in a cell-type and species-specific manner. Hum Gene Ther. 2003;14(12): [18] Van Maele B, De Rijck J, De Clercq E, et al.impact of the central polypurine tract on the kinetics of human immunodeficiency virus type 1 vector transduction. J Virol. 2003;77(8): [19] Chao MV, Rajagopal R, Lee FS. Neurotrophin signalling in health and disease. Clin Sci (Lond). 2006;110(2): [20] Lu P, Tuszynski MH. Growth factors and combinatorial therapies for CNS regeneration. Exp Neurol. 2008;209(2): [21] Zhang M, Wang B, Ni YH, et al. Overexpression of uncoupling protein 4 promotes proliferation and inhibits apoptosis and differentiation of preadipocytes. Life Sci. 2006;79(15): [22] Wen WH, Liu JY, Qin WJ, et al. Targeted inhibition of HBV gene expression by single-chain antibody mediated small interfering RNA delivery. Hepatology. 2007;46(1): [23] Zimmermann TS, Lee AC, Akinc A, et al. RNAi-mediated gene silencing in non-human primates. Nature. 2006;441(7089): [24] Barbin G, Manthorpe M, Varon S. Purification of the chick eye ciliary neuronotrophic factor. J Neurochem. 1984;43(5): [25] Lin LF, Mismer D, Lile JD, et al. Purification, cloning, and expression of ciliary neurotrophic factor (CNTF). Science. 1989;246(4933): [26] Ishii K, Nakamura M, Dai H, et al. Neutralization of ciliary neurotrophic factor reduces astrocyte production from transplanted neural stem cells and promotes regeneration of corticospinal tract fibers in spinal cord injury. J Neurosci Res. 2006;84(8): [27] Lu P, Jones LL, Tuszynski MH. BDNF-expressing marrow stromal cells support extensive axonal growth at sites of spinal cord injury. Exp Neurol. 2005;191(2): [28] Szotek PP, Chang HL, Zhang L, et al. Adult mouse myometrial label-retaining cells divide in response to gonadotropin stimulation. Stem Cells. 2007;25(5): [29] Kato K, Yoshimoto M, Kato K, et al. Characterization of side-population cells in human normal endometrium. Hum Reprod. 2007;22(5): [30] Schwab KE, Gargett CE. Co-expression of two perivascular cell markers isolates mesenchymal stem-like cells from human endometrium. Hum Reprod. 2007;22(11): ISSN CN /R CODEN: ZLKHAH 3609

5 Huang W, et al. Brain-derived neurotrophic factor, ciliary neurotrophic factor and their combination for in vitro differentiation of human... 脑源性神经营养因子和睫状神经营养因子单独或联合体外诱导人脐带血来源的间充质干细胞向神经样细胞分化 * 黄伟 1, 苗宗宁 1, 陈镭 2, 张学光 3 ( 1 无锡市第三人民医院危重病医学科, 江苏省无锡市 ; 2 武警总院脑科系, 天津市 ; 3 苏州大学生物技术研究所, 省重点干细胞实验室, 江苏省苏州市 ) 黄伟, 男,1972 年生, 江苏省无锡市人, 汉族,1995 年湖南医科大学毕业, 硕士, 副主任医师, 主要从事危重病的研究 通讯作者 : 苗宗宁, 副教授, 博士, 无锡市第三人民医院干细胞实验室, 江苏省无锡市 摘要 背景 : 体外培养条件下脐血间充质干细胞可向神经样细胞诱导分化, 一定浓度范围的脑源性神经营养因子和睫状神经营养因子联合体外诱导可获得较高的神经元分化比例 目的 : 观察脑源性神经营养因子和睫状神经营养因子单独或联合体外诱导人脐带血来源的间充质干细胞分化成神经样细胞的可行性 方法 : 取第 5 代的脐血间充质干细胞, 分别用 5,10,20 μg/l 的脑源性神经营养因子和 5,10,20 μg/l 睫状神经营养因子单独或联合诱导脐血源间充质干细胞向神经样细胞分化, 另设空白对照组无任何干预措施 倒置相差显微镜观察细胞形态变化, 于 实验第 1,3,6 天分别进行神经元特异性烯醇化酶和胶质纤维酸性蛋白免疫细胞化学染色, 并计数分化为神经元样细胞及神经胶质细胞的比例 结果与结论 :1 向神经细胞诱导后, 人脐血源间充质干细胞形态明显改变, 胞体收缩, 胞核部分折光性增强, 出现类似于树突及轴突样结构 2 与空白对照组相比, 脑源性神经营养因子和睫状神经营养因子能显著提高人脐血源间充质干细胞分化为神经元的比例 其中 20 μg/l 的脑源性神经营养因子联合 20 μg/l 睫状神经营养因子诱导人脐血源间充质干细胞分化为神经样细胞的比例最高 提示人脐血间充质干细胞经脑源性神经营养因子和睫状神经营养因子体外诱导, 均能够分化为神经样细胞 关键词 : 脑源性神经营养因子 ; 睫状神经营养因子 ; 脐带血 ; 间充质干细胞 ; 诱导 ; 神经样细胞 doi: /j.issn 中图分类号 : R394.2 文献标识码 : A 文章编号 : (2010) 黄伟, 苗宗宁, 陈镭, 张学光. 脑源性神经营养因子和睫状神经营养因子单独或联合体外诱导人脐带血来源的间充质干细胞向神经样细胞分化 [J]. 中国组织工程研究与临床康复, 2010,14(19): [ (Edited by Song LP/Wang L) 来自本文课题的更多信息 -- 基金资助 : 江苏省中医药局计划项目 (HZ07095) 利益冲突 : 无其他利益冲突 ISSN CN /R 2010 年版权归 中国组织工程研究与临床康复 杂志社所有 与干细胞研究相关学术会议介绍 : 本刊学术部 内容简介 网站点击更多 第三届广州国际干细胞与再生生物学论坛, 将于 2010 年 12 月 日在广州举行 本次论坛拟围绕干细胞重编程研究 干细胞的化学生物学研究 胚胎与成体干细胞的应用研究 干细胞与药物研发 发育与模式动物研究 干细胞生物学与克隆等议题展开广泛的交流与讨论, 会议将邀请中科院干细胞与克隆研究海外创新团队成员 发育与生殖重大计划专家组成员 973 首席科学家以及其他国内外本领域知名专家等参会, 与大家共同分享干细胞与再生生物学研究领域的最新进展及成果 探讨干细胞和再生生物学的发展趋势 经中华医学会继续教育部批准, 由空军总医院和中华医学会继续教育部共同主办的 全国造血干细胞移植相关并发症诊治 研讨班, 定于 5 月中旬在北京举行 届时特聘请有关著名专家 教授授课, 主要讲授造血干细胞移植相关并发症如 GVHD 预防与治疗, 肺部并发症 不同时期细菌 病毒 真菌感染诊治等与临床密切相关的课题