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碱性成纤维细胞生长因子刺激下的人胎盘血和脐带血造血祖细胞的体外扩增

Basic fibroblast growth factor-stimulated ex vivo expansion of haematopoietic progenitor cells from human placental and umbilical cord blood.

作者信息

Kashiwakura Ikuo, Takahashi Tsuneo A

机构信息

Department of Radiological Technology, Hirosaki University School of Health Sciences, Hirosaki, Japan.

出版信息

Br J Haematol. 2003 Aug;122(3):479-88. doi: 10.1046/j.1365-2141.2003.04444.x.

Abstract

We investigated whether basic fibroblast growth factor (bFGF) is effective in inducing ex vivo expansion of CD34+ haematopoietic progenitor cells derived from human placental and umbilical cord blood. bFGF significantly promoted the clonal growth of various haematopoietic progenitor cells, including granulocyte-macrophage colony-forming units (CFU-GM), mixed colony-forming units (CFU-Mix) and megakaryocyte colony-forming units (CFU-Meg) under semisolid culture conditions, with an optimal bFGF concentration of 30 ng/ml. CD34+ cells were then cultured in serum-free liquid medium containing various combinations of early-acting cytokines, including thrombopoietin (TPO), stem cell factor (SCF), interleukin 3 (IL-3) and flt3-ligand (FL), with or without bFGF, for 6 and 12 d. Without bFGF, TPO + IL-3, TPO + SCF + FL and TPO +SCF + IL-3 + FL dramatically increased the total numbers of erythroid progenitors, CFU-GM and CFU-Mix by d 12 of culture respectively. However, the addition of bFGF did not promote further proliferation of these progenitors, except for the erythroid progenitors, by d 6 when stimulated with all four cytokines. In contrast, total CFU-Meg numbers were approximately doubled when these cultures were supplemented with bFGF, producing 100- to 120-fold increases compared with the baseline control cultures. These results suggest that bFGF is effective in supporting the generation of megakaryocytic progenitor cells during ex vivo expansion.

摘要

我们研究了碱性成纤维细胞生长因子(bFGF)是否能有效地诱导源自人胎盘和脐带血的CD34+造血祖细胞在体外扩增。在半固体培养条件下,bFGF显著促进了各种造血祖细胞的克隆生长,包括粒细胞-巨噬细胞集落形成单位(CFU-GM)、混合集落形成单位(CFU-Mix)和巨核细胞集落形成单位(CFU-Meg),最佳bFGF浓度为30 ng/ml。然后,将CD34+细胞在含有早期作用细胞因子各种组合的无血清液体培养基中培养,这些细胞因子包括血小板生成素(TPO)、干细胞因子(SCF)、白细胞介素3(IL-3)和fms样酪氨酸激酶3配体(FL),添加或不添加bFGF,培养6天和12天。在没有bFGF的情况下,TPO + IL-3、TPO + SCF + FL和TPO + SCF + IL-3 + FL分别在培养第12天时显著增加了红系祖细胞、CFU-GM和CFU-Mix的总数。然而,在所有四种细胞因子刺激下,到培养第6天时,添加bFGF除了促进红系祖细胞的进一步增殖外,并未促进这些祖细胞的进一步增殖。相反,当这些培养物添加bFGF时,CFU-Meg总数大约增加了一倍,与基线对照培养物相比增加了100至120倍。这些结果表明,bFGF在体外扩增过程中有效地支持了巨核细胞祖细胞的生成。

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