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Neural Regeneration Research 2011年第13期6卷 972-977页

作者： Lei Chen Zhongguo Zhang Bing Chen Xiaozhi Liu Zhenlin Liu Hongliang Liu Gang Li Zhiguo Su Junfei Wang Guozhen Hui Department of Neurosurgery Tianjin Fifth Central Hospital Tianjin 300450 China Department of Colorectal Cancer Surgery Liaoning Provincial CancerHospitail Shenyang 110042 Liaoning Province China Department of Gynaecology and Obstetrics Shanghai Pudong New District People's Hospitall Shanghai 201200 China Department of Neurosurgery First Affiliated Hospital of Soochow University Suzhou 215006 Jiangsu Province 'China

human umbilical cord blood was collected from full-term deliveries scheduled for cesarean section. Mononuclear cells were isolated, amplified and induced as mesenchymal stem cells. Isolated mesenchymal stem cells tested positive for the marker CD29, CD44 and CD105 and negative for typical hematopoietic and endothelial markers. Following treatment with neural induction medium containing brain-derived neurotrophic factor for 7 days, the adherent cells exhibited neuron-like cellular morphology. Immunohistochemical staining and reverse transcription-PCR revealed that the induced mesenchymal stem cells expressed the markers for neuron-specific enolase and neurofilament. The results demonstrated that human umbilical cord blood-derived mesenchymal stem cells can differentiate into neuron-like cells induced by brain-derived neurotrophic factor in vitro.

关键词： human umbilical cord blood purification and culture brain-derived neurotrophic factor neuron-like cells neural regeneration

Characterization of Adherent Nonhematopoietic Cells Derived from human umbilical cord blood

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Tsinghua Science and Technology 2003年第4期8卷 412-416页

作者：安小惠蔡国平 Department of Biological Sciences and Technology Tsinghua University Beijing 100084 China

To confirm and characterize the adherent fibroblast like progenitors in human umbilical cord blood, we isolated mononuclear cells from human umbilical cord blood by Ficoll Hypaque. Two main morphologically different kinds of cells were formed by culturing the cells in collagen coated 24 well plastic dishes and flasks. One type was the adherent fibroblast like cells, while the other was loosely adherent clonally expanded round cells. Our experiments demonstrate that the adherent fibroblast like cells possess multilineage potential, including the ability to differentiate into endothelial like cells and to express the mesenchymal cell marker.

关键词： human umbilical cord blood mesenchymal cells endothelial cells

Experimental study on ex vivo expanded hematopoietic stem/progenitor in the two step culture from human umbilical cord blood transplanted into NOD/SCID mice

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Frontiers in Biology 2006年第2期1卷 137-141页

作者： Jia Bingbing Xiang Ying Xie Chungang Wang Jinfu College of life Sciences Zhejiang UniversityHangzhou 310058China

The effects of hematopoietic stem/progenitor cells(HSPCs)expanded in the two step coculture with human bone marrow mesenchymal stem cells(hMSCs)on the hematopoietic reconstruction of irradiated NOD/SCID mice were *** cells(MNCs)were isolated from human umbilical cord blood(UCB)and cultured in the non-coculture scheme of rhSCF+rhG−CSF+rhMDGF combination and the coculture scheme of rhSCF+rhG−CSF+rhMDGF+***-irradiated NOD/SCID mice were transplanted with ex vivo expanded HSPCs with the dose of 8.5×10^(6) cells per *** transplantation,the dynamics of WBC in the transplanted mice was measured periodically,and the Alu sequence fragment special for human in the transplanted mice was inspected by *** showed that the coculture scheme increased proliferation of UCB-derived *** transplantation with expanded HSPCs,the population of WBC in the transplanted mice increased in 12 d and reached the first peak in 25 d,then showed the second increasing of WBC in 45~55 *** cells from the coculture scheme appeared to be favorable for the second increasing of WBC in the transplanted *** 85 d,the Alu sequence fragment was detected in the probability of 87.5%(7/8)for the non-coculture scheme and 88.9%(8/9)for the coculture scheme.

关键词： human umbilical cord blood hematopoietic stem/progenitor cells ex vivo expansion NOD/SCID mice transplantation

Multiorgan engraftment of donor-derived human cells in mice by transplanting human umbilical cord blood-derived CD133(+) cells into murine blastocysts

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Multiorgan engraftment of donor-derived human cells in mice ...

human umbilical cord-derived endothelial progenitor cells promote growth cytokines-mediated neorevascularization in rat myocardial infarction

第九届中国实验动物科学年会（2010新疆）

作者： Yan Sun~2,Dong Xiao~1 1) Center of Experimental Animals,Sun Yat-sen University,Guangzhou 510080,Guangdong Province, China 2) Cancer Research Institute,Southern Medical University,Guangzhou 510515,Guangdong Province, China

Objective A relatively unperturbed developmental system in vivo was employed to fully evaluate the plasticity of human CD133(+) stem ***:To address this,we transplanted human umbilical cord blood(hUCB)-derived CD133(+) cells into murine blastocysts to develop the noninjury human-mouse chimeras(HMCs),in which the donor-derived human cell engraftment in multiple organs of recipients was detected by human-specific PCR for human Alu gene,in situ hybridization(ISH) for human Alu sequences and immunohistochemistry(IHC).Results:we took advantage of the proliferative and permissive environment of the developing pre-immune embryo to successfully develop HMCs,in which blastocyst transplantation of human CD133(+) stem cell population led to the donor-derived human cell engraftment in multiple organs(i.e.,liver,spleen,thymus,heart,kidney,blood,lung,gut,etc),as revealed by PCR,ISH and IHC for detecting humanβ2-microglobulin(β2-M) *** data demonstrated the formation of donor-derived human hepatocyte-like cells(hHLCs) with different cellular phenotypes,as demonstrated by positively immunostaining for human-specific alpha-fetoprotein(AFP),cytokeratin 19(CK19),cytokeratin 8(CK8) and cytokeratin 18(CK18) in the recipient ***,the transplanted human donor cells underwent site-specific differentiation into vWF-positive, CD34-positive and vimentin-positive human endothelial-like cells within various blood vessel(i.e,arteries and veins) walls of the multiple organs in ***:Taken together,these in vivo findings strongly indicate the plasticity of CD133(+) stem cells in ***133(+) stem cells might be useful for research and for clinical application.

关键词： human umbilical cord blood CD133(+) cells Mice Blastocyst microinjection Embryonic microenvironment Plasticity Chimerism human-mouse chimeras(HMCs) humanized liver human hepatocyte-like cells human endothelial-like cells Stem cells

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Chinese Medical Journal 2009年第5期122卷 548-555页

作者： HU Cheng-heng LI Zhi-ming DU Zhi-min ZHANG Ai-xia YANG Da-ya WU Gui-fu Division of Cardiology First Affiliated Hospital and Key Laboratory on Assisted Circulation Ministry of Health of China Sun Yat-sen University Guangzhou Guangdong 510080 China

Background Cell-based vascular therapies of endothelial progenitor cells （EPCs） mediated neovascularization is still a novel but promising approach for the treatment of ischemic disease. The present study was designed to investigate the therapeutic potentials of human umbilical cord blood-derived EPCs （hUCB-EPCs） in rat with acute myocardial infarction. Methods human umbilical cord blood （hUCB） mononuclear cells were isolated using density gradient centrifugation from the fresh human umbilical cord in healthy delivery woman, and cultured in M199 medium for 7 days. The EPCs were identified by double-positive staining with 1,1＇-dioctadecyl-3,3,3＇,3＇-tetramethylindocarbocyanine percholorate-labeled acetylated low-density lipoprotein （DiI-Ac-LDL） and fluorescein isothiocyanate-conjugated Ulex europaeus lectin （FITC-UEA-I）. The rat acute myocardial infarction model was established by the ligation of the left anterior descending artery. The hUCB-EPCs were intramyocardially injected into the peri-infarct area. Four weeks later, left ventricular function was assessed by a pressure-volume catheter. The average capillary density （CAD） was evaluated by anti-VIII immunohistochemistry staining to reflect the development of neovascularization at the peri-infarct area. The graft cells were identified by double immunofluorescence staining with human nuclear antigen （HNA） and CD31 antibody, representing human origin of EPCs and vascular endothelium, respectively. Expressions of cytokines, proliferating cell nuclear angigen （PCNA）, platelet endothelial cell adhesion molecule （PECAM） and vascular endothelial growth factor （VEGF） were detected to investigate the underlying mechanisms of cell differentiation and revascularization. Results The donor EPCs were detectable and integrated into the host myocardium as confirmed by double-positive immunofluorescence staining with HNA and CD31. And the anti-VIII staining demonstrated a higher degree of microvessel formation in EPCs transplanted

关键词： human umbilical cord blood endothelial progenitor cells acute myocardial infarction neovascularization

A secretory function of human insulin-producing cells in vivo

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Hepatobiliary & Pancreatic Diseases International 2009年第3期8卷 255-260页

作者： Hu, Yan-Hua Wu, De-Quan Gao, Feng Li, Guo-Dong Yao, Lei Zhang, Xin-Chen Harbin Med Coll Dept Gen Surg Affiliated Hosp 2 Harbin 150086 Peoples R China

BACKGROUND: Mesenchymal stem cells derived from human umbilical cord blood (UCB-MSCs) have good research and application prospects in the treatment of diabetes. We once induced UCB-MSCs to differentiate into insulin-producing cells (IPCs) in vitro, but we did not know the functions of these cells in vivo. The aim of this study was to assess the functional effects of IPCs on insulin secretion and their role in the treatment of diabetes in vivo. METHODS: UCB-MSCs were induced to IPCs by an inducing protocol with extracellular matrix gel. BALB/C nude mice were made hyperglycemic by intraperitoneal injection of streptozotocin. The diabetic mice were transplanted with 1x10(7) IPCs under the renal capsule or with phosphate-buffered saline as a control. After transplantation, the grafts were analyzed by immunocytochemistry for the expression of human insulin; the serum human insulin levels were measured; and blood glucose and body weight status were monitored. RESULTS: Immunofluorescence showed that numerous IPCs under the kidney capsule were insulin-positive. On day 14 after transplantation, the serum human insulin level of the treatment group (n=9) averaged 0.44 +/- 0.12 mU/L, which was higher than that of the control group (n=9) that did not express insulin (t=10.842, Phuman insulin in diabetic mice, but more research is needed to make further use of them to regulate hyperglycemia and body weight in vivo. (Hepatobiliary Pancreat Dis Int 2009; 8: 255-260)

关键词： human umbilical cord blood mesenchymal stem cells insulin-producing cells diabetes cell transplantation