CD34+ cell expansion and expression of lineage markers during liquid culture of human progenitor cells
CD34+ cell expansion and expression of lineage markers during liquid culture of human progenitor cells
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人类祖细胞液体培养过程中 CD34+ 细胞的扩增和谱系标记物的表达
DOI：
10.1002/stem.5530130208
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发表时间：
1995
期刊：
影响因子：
5.2
通讯作者：
J. Cone
中科院分区：
文献类型：
--
作者：
M. Warren;W. L. Rose;L. Beall;J. Cone
建立了一套基于96孔的人造血祖细胞悬浮培养系统，用于监测CD34+细胞向特定谱系的迁移和分化，以及CD34+细胞的维持和体外扩增。采用酶联免疫吸附试验(EL ISA)检测细胞表面成熟标志和谱系标志的表达。从脐血和胎肝(纯度90%)中分离CD34+细胞，在96孔板中液体培养10天。然后用戊二醛-多聚甲醛混合物固定细胞，将细胞牢固地附着在塑料上。使用针对细胞表面标志物的适当一抗进行了ELISA法。检测CD14(单核细胞)、CD15(中性粒细胞)、血小板膜糖蛋白(GP)IIb/IIIa(CD41a，巨核细胞)和血糖蛋白A(红系)的表达。白细胞介素3(IL-3)和干细胞因子(SCF)联合应用刺激CD14、CD15和GP IIb/IIIa的表达。血统限制性生长因子如促红细胞生成素(EPO)与干细胞因子(SCF)联合刺激血糖素A的表达。IL-3、SCF和EPO三因素联合刺激所有四种血统标志物的表达。通过酶联免疫吸附试验和流式细胞仪检测，其他多种生长因子组合都能刺激髓系和巨核细胞的生长，但只有当EPO包含在生长因子混合物中时，红系生长才会出现。在无血清或有血浆的培养液中，CD14的表达显著降低，而血糖素A的表达在无血清的培养液中显著升高。随着培养时间的推移，CD34的表达也被测量到，虽然很低，但可以检测到，并在培养期间略有增加。CD34在胎肝细胞中的表达普遍高于脐血细胞。本研究证明，用酶联免疫吸附试验检测CD34和谱系标记物是一种客观的、高容量的表征祖细胞生长的方法。
A 96‐well‐based suspension culture system for human hematopoietic progenitor cells has been developed to monitor the commitment and differentiation of CD34+ cells to specific lineages and the maintenance and expansion of CD34+ cells in vitro. The expression of maturation and lineage markers on the cells in culture was measured by enzyme‐linked immunosorbent assay (ELISA). CD34+ cells were isolated from umbilical cord blood and fetal liver (90% purity) and were grown in liquid culture in 96‐well plates for 10 days. The cells were then fixed with a glutaraldehyde‐paraformaldehyde mixture, attaching the cells firmly to the plastic. An ELISA was performed, using appropriate primary antibodies directed against cell surface markers. The expression of four different lineage markers was measured: CD14 (monocyte), CD15 (neutrophil), platelet glycoprotein (GP) IIb/IIIa (CD41a, megakaryocyte) and glycophorin A (erythroid). The two‐growth factor combination of interleukin 3 (IL‐3) and stem cell factor (SCF) stimulated expression of CD14, CD15 and GP IIb/IIIa. Lineage‐restricted growth factors such as erythropoietin (EPO), in combination with SCF, stimulated expression of glycophorin A. The three‐factor combination of IL‐3, SCF and EPO stimulated expression of all four lineage markers. Other multiple growth factor combinations all stimulated myeloid and megakaryocyte growth, as measured by ELISA and flow cytometry, but erythroid growth was present only when EPO was included in the growth factor mixture. In serum‐free medium or plasma‐containing medium, CD14 expression was markedly reduced, whereas glycophorin A expression was greatly elevated in serum‐free medium. The expression of CD34 was also measured over time in culture and was low but detectable and slightly increased over the culture period. CD34 expression was generally higher with fetal liver cells than with cord blood cells. This study demonstrates the detection of CD34 and lineage markers by ELISA as an objective, high‐capacity method of characterizing progenitor cell growth.
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影响因子：
56.9
作者：
WRIGHT, SD;RAMOS, RA;MATHISON, JC
通讯作者：
MATHISON, JC
影响因子：
2.6
作者：
Civin,CI
通讯作者：
Civin,CI
使用白介素 1 加试剂盒配体离体扩增骨髓进行骨髓移植可加速小鼠造血重建，而不会损失干细胞谱系和增殖潜力。
影响因子：
20.3
作者：
Muench,MO;Firpo,MT;Moore,MA
通讯作者：
Moore,MA
影响因子：
20.3
作者：
H. Mayani;W. Dragowska;P. Lansdorp
通讯作者：
H. Mayani;W. Dragowska;P. Lansdorp
造血调节因子：冗余还是微妙？
影响因子：
20.3
作者：
Metcalf,D
通讯作者：
Metcalf,D