期刊文献+

利用外周血单个核细胞生产成熟红细胞 被引量:2

Production of Mature Red Blood Cell by Using Peripheral Blood Mononuclear Cells
下载PDF
导出
摘要 体外制备红细胞的方法大多使用脐血、骨髓或外周动员的CD34+细胞或胚胎干细胞作为起始材料。本研究利用外周血单个核细胞为原料在体外生产成熟红细胞。以血液滤涂白细胞后剩余的白膜层为原料,从中分离出单个核细胞,然后通过4步培养法制备成熟的红细胞。结果表明:经过不同细胞因子组合的诱导以及小鼠基质细胞系MS-5的支持培养,外周血单个核细胞的扩增倍数达到约1 000倍,其中约90%的细胞是与正常红细胞形态相似的无核成熟红细胞。集落形成能力分析显示,本研究培养体系可使单个核细胞中的祖细胞增殖并且只向红系方向分化。结构和功能分析结果表明,体外培养出的红细胞的平均细胞体积(MCV)为118±4 fl,比正常红细胞(80-100 fl)稍大。平均血红蛋白含量(MCH)为36±1.2 pg,比正常的红细胞水平(27-31 pg)稍高。红细胞的最大变形指数(DImax)为0.46,接近正常红细胞水平。葡萄糖6磷酸脱氢酶及丙酮酸激酶的含量与年轻红细胞的特性一致。结论:利用外周血白膜层中含有的造血干祖细胞,通过体外培养能够生产出成熟红细胞,它是一种取材方便、成本低廉的红细胞生产原料,本研究培养体系适于利用单个核细胞生产成熟红细胞。 Most protocols for in vitro producing red blood cells(RBC) use the CD34 + cells or embryonic stem cells from cord blood,bone marrow or peripheral blood as the start materials.This study was purposed to produce the mature RBC in vitro by using peripheral blood mononuclear cells as start material.The peripheral blood mononuclear cells (PBMNC) were isolated from buffy coat after blood leukapheresis,the mature red blood ceils(RBC) were prepared by a 4-step culture protocol.The results showed that after culture by inducing with the different sets of cytokines and supporting by mouse MS-5 cell line,the expansion of PBMNC reached about 1000 folds at the end of the culture.About 90% of cultured RBC were enucleated mature cells which had the comparable morphological characteristics with normal RBC.Colony-forming assays showed that this culture system could stimulate the proliferation of progenitors in PBMNC and differentiate into erythroid cells.The structure and function analysis indicated that the mean cell volume of in vitro cultured RBC was 118 ±4 fl,which was slight larger than that of normal RBC (80-100 fl) ; the mean cell hemoglobin was 36 ± 1.2 pg,which was slight higher than that of normal RBC (27-31 pg) ; the maximal deformation index was 0.46,which approachs level of normal RBC; the glucose-6-phosphate dehydrogenase and pyrurvate kinase levels was consistant with young RBC.It is concluded that PBMNC are feasble,convenient and low-cost source for producing cultured RBC and this culture system is suitable to generate the RBC from PBMNC.
出处 《中国实验血液学杂志》 CAS CSCD 北大核心 2014年第5期1435-1441,共7页 Journal of Experimental Hematology
关键词 外周血单个核细胞 体外培养 红细胞 造血干细胞 peripheral blood mononuclear cell in vitro culture RBC hematopoietic stem cell
  • 相关文献

参考文献14

  • 1Neildez-Nguyen TM,Wajcman H,Marden MC,et al.Human erythroid cells produced ex vivo at large scale differentiate into red blood cells in vivo.Nat Biotechnol.2002;20(5):467-472. 被引量:1
  • 2Giarratana MC,Kobari L,Lapillonne H,et al.Ex vivo generation of fully mature human red blood cells from hematopoietic stem cells.Nat.Biotechnol.2005;23(1):69-74. 被引量:1
  • 3Ronzoni L,Bonara P,Rusconi D,et al.Erythroid differentiation and maturation from peripheral CD34+cells in liquid culture:Cellular and molecular characterization.Blood Cells,Molecules,and Diseases,2008;40(2):148-155. 被引量:1
  • 4Fujimi A,Matsunaga T,Kobune M,et al.Ex vivo large-scale generation of human red blood cells from cord blood CD34+cells by co-culturing with macrophages.Int J Hematol.2008;87:339-350. 被引量:1
  • 5Baek EJ,Kim HS,Kim S,et al.In vitro clinical-grade generation of red blood cells from human umbilical cord blood CD34+cells.Transfusion,2008;48(10):2235-2245. 被引量:1
  • 6贾延军,刘江,张可莹,单小燕,李伟,何晓梅,王立君,刘娜,王琳,崔爽,倪雷,赵博涛,龚志尹,王东玫,高颂明,张志欣.脐血CD34^+细胞体外诱导扩增红系细胞的探索[J].中国实验血液学杂志,2009,17(3):787-792. 被引量:2
  • 7贾延军,刘江,张可莹,单小燕,李伟,何晓梅,王立君,刘娜,王琳,崔爽,倪雷,赵博涛,龚志尹,王东玫,高颂明,张志欣.培养条件对CD34^+细胞体外诱导分化和扩增红细胞的影响[J].中国输血杂志,2010,23(2):107-110. 被引量:5
  • 8Lu SJ,Feng Q,Park JS,et al.Biologic properties and enucleation of red blood cells from human embryonic stem cells.Blood,2008;112(12):4475-4484. 被引量:1
  • 9Boehm D,Murphy WG and Al-Rubeai M.The potential of human peripheral blood derived CD34+cells for ex vivo red blood cell production.J Biotechnol,2009;144(2):127-134. 被引量:1
  • 10Qiu CH,Olivier EN,Velho M,Bouhassira EE.Globin switches in yolk sac primitive and fetal-like definitive red blood cells produced from human embryonic stem cells.Blood,2008;111(4):2400-2408. 被引量:1

二级参考文献19

  • 1Ballen KK.New trends in umbilical cord blood transplantation.Blood,2005 , 105:3786 -3792 被引量:1
  • 2Munugalavadia V,Kapur R.Role of c-kit and erythropoietin receptor in erythropoiesis.Crit Rev Oncol Hematol,2005 , 54:63 -75 被引量:1
  • 3Okumura N,Tsuji K,Nakahata T.Changes in celd surface antigen expressions during proliferation and differentiation of human erythroid progenitors.Blood,1992 , 80:642-650 被引量:1
  • 4Zhu Y,D 'Andrea AD.The molecular physiology of erythropoietin and the erythropoietin receptor.Curr Opin Hematol,1994, 1:113-118 被引量:1
  • 5Wang J,Tang ZY,Ka W,et al.Synergistic effect of cytokines EPO,IL-3 and SCF on the proliferation,differentiation and apoptosis of erythroid progenitor cells.Clin Hemorheol Microcirc,2007 ,37:291-199 被引量:1
  • 6Hoffman R,Tong J,Brandt J,et al.The in vitro and in vivo effects of stem cell factor on human hematopoiesis.Stem Cells,1993, 11(suppl 2):76-82 被引量:1
  • 7Giarratana MC,Kobari L,Lapillonne H,et al.Ex vivo generation of fully mature human red blood cells from hematopoietic stem cells.Nat Biotechnol,2005 , 23:69-74 被引量:1
  • 8Miharada K,Hiroyama T,Sudo K,et al.Efficient enucleation of erythroblast differentiated in vitro from hematopoietic stem and progeniter cells.Nat Biotechnol,2006, 24:1255-1256 被引量:1
  • 9Kawano Y,Kohune M,Yamaguchi M,et al.Ex vivo expansion of human umblilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit (hTERT) -transfected human stromal cells.Blood,2003, 101:532 -540 被引量:1
  • 10Miharada K,Hiroyama T,Sudo K,et al.Lipocalin 2 functions as a negative regulator of red blood cell production in an autocrine fashion.FASEB J,2005, 19:1881-1883 被引量:1

共引文献4

同被引文献7

引证文献2

二级引证文献1

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部