期刊文献+

间充质干细胞及其培养上清与融冻产物对脐血CD34^+细胞扩增及分化的作用 被引量:2

Effects of Mesenchymal Stem Cells Feeder Layer, Culture Sera and Freeze-thaw Lysates on Expansion and Differentiation of Cord Blood CD34^+ Cells in Vitro
下载PDF
导出
摘要 【目的】探讨人骨髓间充质干细胞(MSC)及其培养上清、融冻产物对脐血CD34+细胞扩增及分化的作用。【方法】分离纯化人骨髓MSC,收集纯化三代的MSC培养上清,采用反复冻融法获得MSC融冻产物,采用免疫磁珠分选系统纯化脐血CD34+细胞。实验分为4组,分别为含MSC滋养层组、MSC上清组、MSC融冻产物组及单纯造血生长因子(HGFs)对照组。流式细胞仪检测人脐血CD34+细胞在这四种培养体系中第6天和第12天有核细胞数及CD34+、CD34+CD38-、CD41+、CD19+、CD3+细胞含量。【结果】①MSC滋养层组对脐血有核细胞和对脐血CD34+、CD34+CD38-的扩增能力最强,MSC培养上清组和MSC滋养层相比具有类似的作用,但其作用强度减弱(P<0.05)。②MSC培养上清和MSC均具有抑制脐血CD34+细胞分化为CD3+和CD19+细胞的作用,差异无显著性(P>0.05)。③MSC培养上清和MSC均具有促进脐血CD34+细胞向CD41+细胞分化的作用,MSC滋养层的作用强于MSC培养上清(P<0.05)。④MSC融冻产物已完全丧失对脐血CD34+细胞扩增和分化作用,与单纯因子对照组相似(P>0.05)。【结论】MSC培养上清对脐血CD34+、CD34+CD38-细胞均具有扩增作用,而且可促进脐血CD34+细胞向CD41+细胞分化。 【Objective】 To investigate the effects of mesenchymal stem cells (MSC) feeder layer,culture sera and freeze- thaw lysates on expansion and differentiation of cord blood CD34+ cells in vitro.【Methods】 MSC were isolated from human bone marrow and cultured until the third passage.Sera were obtained from the cultured MSC,and freeze-thaw lysates were obtained by repeated freeze-thaw procedures.Cord blood CD34+ cells were isolated by magnetic cell separation system,and were co-cultured with the MSC feeder layer,culture sera,freeze-thaw lysates and hematopoietic growth factors (HGFs),respectively.The nucleated cells,CD34+ cells,CD34+ CD38- cells,CD41+ cells and CD3+ cells in the above culture system were detected by flow cytometry on day 6 and day 12.【Results】 ①MSC feeder layer had a strong effect on nucleated cells,CD34+,CD34+ CD38- cells expansion.The MSC sera and freeze-thaw lysates had similar effect on cell expansion,but the effect was weaker than that of feeder layer (P 0.05).②Both MSC sera and feeder layer inhibited cord blood CD34+ cells differentiation toward CD3+ cells or CD19+ cells,and no significant differences were found between these two groups (P 0.05).③Both MSC sera and feeder layer promoted cord blood CD34+ cells differentiation toward CD41+ cells,and the effect was stronger in the feeder layer than that of the sera (P 0.05).④Freeze-thaw lysates had no effect on cell expansion and differentiation,and were similar with that of HGFs (P 0.05).【Conclusions】 The MSC sera have positive effects on expansion of cord blood CD34+ and CD34+ CD38- cells,moreover they have the ability of promoting cord blood CD34+ cells differentiation toward CD41+ cells.
出处 《中山大学学报(医学科学版)》 CAS CSCD 北大核心 2010年第1期39-44,共6页 Journal of Sun Yat-Sen University:Medical Sciences
基金 国家自然科学基金(30872786) 广东省科技计划引导项目(2008B030301305)
关键词 间充质干细胞 脐血CD34+细胞 MSC培养上清 MSC融冻产物 mesenchymal stem cells MSC culture sera MSC freeze-thaw lysates CD34+ cells of cord blood
  • 引文网络
  • 相关文献

参考文献15

  • 1Ball LM,Bemardo ME,Locatelli F,et al.Potential role of mesenchymal stromal cells in pediatric hematopoietic SCT[J].Bone Marrow Transplant,2008,42 (Suppl 2):S60-66.
  • 2Polchert D,Sobinsky J,Douglas G,et al.IFN-gamma activation of mesenchymal stem cells for treatment and prevention of graft versus host disease[J].Eur J Immunol,2008,38(6):1479-1482.
  • 3Le Blanc K,Frassoni F,Ball L,et al.Mesenchymal stem cells for treatment of steroid-resistant,severe,acute graft-versus-host disease:a phase Ⅱ study[J].Lancet,2008,371 (9624):1553-1554.
  • 4del Caf(n)izo MC,Lopez N,Vazquez L,et al.Hematopoietic damage prior to PBSCT and its influence on hematopoietic recovery[J].Haematologica,1999,84(6):511-516.
  • 5Cui J,Wahl RL,Shen T,et al.Bone marrow cell trafficking following intravenous administration[J].Br J Haematol,1999,107(4):895-902.
  • 6Huang GP,Pan ZJ,Jia BB,et al.Ex vivo expansion and transplantation of hematopoietic stem/progenitor cells supported by mesenchymal stem cells from human umbilical cord blood[J].Cell Transplant,2007,16 (6):579-585.
  • 7Bakhshi T,Zabriskie RC,Bodie S,et al.Mesenchymal stem cells from the Wharton's jelly of umbilical cord segments provide stromal support for the maintenance of cord blood hematopoietic stem cells during long-term ex vivo culture[J].Transfusion,2008,48(12):2638-2644.
  • 8Isaikina Y,Shman T.Influence of mesenchymal stem cells derived from bone marrow of children with oncohematological diseases on proliferation and selfrenewal of hematopoietic progenitor cells in vitro[J].Exp Oncol,2008,30 (2):121-128.
  • 9Tian Y,Deng YB,Huang Y J,et al.Bone marrowderived mesenchymal stem cells decrease acute graftversus-host disease after aliogeneic hematopoietic stem cells transplantation[J].Immunol Invest,2008,37 (1):29-42.
  • 10周敦华,魏菁,黄绍良,方建培,包蓉,吴燕峰,黎阳.人脐血来源MSCs的主要生物学特性的研究[J].中国病理生理杂志,2006,22(8):1606-1609. 被引量:7

二级参考文献21

  • 1Erices A,Conget PA,Minguell JJ.Mesenchymal progenitor cells in human umbilical cord blood[J].Br J Haematol,2000,109(1):235 -242.
  • 2Bieback K,Kern S,Kluter H,et al.Critical parameters for the isolation of mesenchymal stem cells from umbilical cord blood[J].Stem Cells,2004,22(4):625-634.
  • 3Lee KD,Kuo TK,Whang-Peng J,et al.In vitro hepatic differentiation of human mesenchymal stem cells[J].Hepatology,2004,40(6):1275-1284.
  • 4Gang EJ,Hong SH,Jeong JA,et al.In vitro mesengenic potential of human umbilical cord blood-derived mesenchymal stem cells[J].Biochem Biophys Res Commun,2004,321(1):102 -108.
  • 5Bianco P,Riminucci M,Gronthos S,et al.Bone marrow stromal stem cells:nature,biology,and potential applications[J].Stem Cells,2001,19(3):180-192.
  • 6Dovovan PJ,Gearheart J.The end of the beginning for pluripotent stem cells[J].Nature,2001,414(2):92 -96.
  • 7Wang JF,Wang LJ,Wu YF,et al.Mesenchymal stem/progenitor cells in human umbilical cord blood as support for ex vivo expansion of CD34 (+) hematopoietic stem cells and for chondrogenic differentiation[J].Haematologica,2004,89(7):837-844.
  • 8姜学英 苏月来 杨少光等.人骨髓基质细胞系的建立、生物学特征和功能研究[J].中华血液学杂志,1988,(1):661-664.
  • 9Majumdar MK,Thiede MA,Haynesworth SE,et al.Human marrow-derived mesenchymal stem cells (MSCs) express hematopoietic cytokines and support long-term hematopoiesis when differentiated toward stromal and osteogenic lineages.J Hematother Stem Cell Res,2000; 9:841-848
  • 10Friedenstein AJ,Gorskaja JF,Kulaqina NN.Fibroblast precursors in normal and irradiated mouse hematopoietic organs.Exp Hematol,1996; 4:267-274

共引文献11

同被引文献34

  • 1蔡耘,黄绍良,陈凤英,黄文革,魏菁,车丽洪.同种异基因小鼠骨髓腔内脐血移植模型的建立及其对造血干细胞植入的影响[J].中山大学学报(医学科学版),2005,26(6):644-650. 被引量:8
  • 2Angelopoulou M,Novelli E,Grove JE,et al.Cotransplantation of human mesenchymal stem cells enhances human myelopoiesis and megakaryocytopoiesis in NOD/SCID mice.Exp Hematol.2003;31(5):413-420.
  • 3Deans RJ,Moselay AB.Mesenchymal stem cells:biology and potential clinical uses.Exp Hematol 2000;28(8):875-884.
  • 4Le Blanc K,Ringden O.Mesenchymal stem cells:properties and role in clinical bone marrow transplantation.Cur Opin Immunol 2006;18(5):586-591.
  • 5Lu LL,Liu YJ,Yang SG,et al.lsolation and characterization of human umbilical cord mesenchymal stem cells with hematopoiesis-supportive function and other potentials.Haematologica 2006;91(8):1017-1026.
  • 6D'lppolito G,Schiller PC,Ricordi C,et al.Age-related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow.J Bone Miner Res 1999;14(7):1115-1122.
  • 7D'lppolito G,Howard GA,Roos BA,et al.Sustained stromal stem cell self-renewal and osteoblastic difrerentiation during aging.Rejuv Res.2006;9(1):10-19.
  • 8Mareschi K,Ferrero I,Rustichelli D,et al.Expansion of mesenchymal stem cells isolated from pediatric and adult donor bone marrow.J Cell Biochem.2006;97(4):744-754.
  • 9Ming-song Tsai,Shiaw-Min Hwang,Kuang-Den Chen,et al.Functional network analysis of the transcriptomes of mesenchymal stem cells derived from amniotic fluid,amniotic membrane,cord blood and bone marrow.Stem Cell.2007;25(10):2511-2523.
  • 10Parolini O,Alviano F,Bagnara GP,et al.Concise review:isolation and characterization of cells from human term placenta:outcome of the first international Workshop on Placenta Derived Stem Cells Stem Cells 2008;26(2):300-311.

引证文献2

二级引证文献5

;
使用帮助 返回顶部