Proteomic Profiling of Ex Vivo Expanded CD34-Positive Haematopoetic Cells Derived from Umbilical Cord Blood
Proteomic Profiling of Ex Vivo Expanded CD34-Positive Haematopoetic Cells Derived from Umbilical Cord Blood
复制标题
DOI：
10.1155/2013/245695
复制
发表时间：
2013-01-01
影响因子：
4.3
通讯作者：
Stuehler, Kai
中科院分区：
文献类型：
--
作者：
Falkenberg, Heiner;Radke, Teja Falk;Stuehler, Kai
通过应用特定的细胞因子体外扩增造血细胞是克服脐血移植中造血干细胞数量有限的障碍的一种方法。虽然许多方案描述了细胞总数和CD34阳性细胞数量的有效增加，但仍不清楚这一过程是否以及如何实际影响细胞的特性。在所发表的文献中，从脐带血中分离出CD34阳性细胞，并在添加干细胞因子(SCF)、血小板生成素(THPO)、白介素6(IL-6)和FMS相关酪氨酸激酶3配体(Flt3lg)的培养液中扩增长达7天。分别于第3、7天收集扩增细胞，用流式细胞仪和定量蛋白质组学方法进行分析。共鉴定出2970个蛋白质，蛋白质组学分析显示其中440个蛋白质在体外扩增过程中含量发生了显著变化。尽管3d后造血细胞表面仍有CD34的表达，但蛋白质谱发生了较大的变化，而进一步扩增对蛋白质组水平的影响较小。生物学过程的浓缩分析清楚地表明，在表达的头三天内，蛋白质组就已经向蛋白质生物合成表型转变。
Ex vivo expansion of haematopoetic cells by application of specific cytokines is one approach to overcome boundaries in cord blood transplantation due to limited numbers of haematopoetic stem cells. While many protocols describe an effective increase of total cell numbers and the amount of CD34-positive cells, it still remains unclear if and how the procedure actually affects the cells' properties. In the presented publications, CD34-positive cells were isolated from cord blood and expanded for up to 7 days in media supplemented with stem cell factor (SCF), thrombopoietin (THPO), interleukin 6 (IL-6), and fms-related tyrosine kinase 3 ligand (FLT3lg). At days 3 and 7, expanded cells were harvested and analyzed by flow cytometry and quantitative proteomics. 2970 proteins were identified, whereof proteomic analysis showed 440 proteins significantly changed in abundance during ex vivo expansion. Despite the fact that haematopoetic cells still expressed CD34 on the surface after 3 days, major changes in regard to the protein profile were observed, while further expansion showed less effect on the proteome level. Enrichment analysis of biological processes clearly showed a proteomic change toward a protein biosynthesis phenotype already within the first three days of expression.