Shahrabi Saeid, Kaviani Saeid, Soleimani Masoud, Pourfathollah Ali Akbar, Bakhshandeh Behnaz, Hajizamani Saeideh, Saki Najmaldin
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran; Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Iran J Med Sci. 2017 Jan;42(1):40-47.
Human umbilical cord blood (HUCB) is an acceptable and readily accessible source of stem cells. There is an ongoing interest in cord blood stem cell therapies; however, little is known about the possible unfavorable effects of laboratory modifications on the isolated HUCB cells. The involvement of miRNAs in several biological processes has been shown. The aim of this study was to evaluate the possible changes in miRNA expression profiles in CD133 hematopoietic cells after culture.
HUCBCD133 hematopoietic stem cells were isolated by magnetic-activated cell sorting, and then the cells were counted using flow cytometry. The cells were divided into 2 groups. In the first group, RNA was extracted and the cells of the second group were cultured for 12 days and then these cells were used to assay miRNAs expression using real-time qPCR.
The results showed that the expression of 349 out of 1,151 screened miRNAs was upregulated following a 12-day culture of CD133 cells, whereas the expression of 293 miRNAs was downregulated. In addition, the expression of 509 miRNAs was not significantly altered. Another analysis involving the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to the selected miRNAs was also conducted.
Based on our results, the expansion of HUCB resulted in altered expression levels of miRNAs. This study provides information on the effects of 2-dimensional culture of hematopoietic cells prior to transplantation for more successful transplantation.
人脐带血（HUCB）是一种可接受且易于获取的干细胞来源。人们对脐带血干细胞疗法一直很感兴趣；然而，对于实验室操作对分离出的HUCB细胞可能产生的不利影响知之甚少。已有研究表明miRNA参与了多个生物学过程。本研究的目的是评估培养后CD133造血细胞中miRNA表达谱的可能变化。
通过磁珠分选法分离HUCB CD133造血干细胞，然后使用流式细胞术对细胞进行计数。将细胞分为两组。第一组提取RNA，第二组细胞培养12天，然后使用实时定量PCR检测这些细胞的miRNA表达。
结果显示，在对CD133细胞进行12天培养后，1151个筛选出的miRNA中有349个的表达上调，而293个miRNA的表达下调。此外，509个miRNA的表达没有显著变化。还对与所选miRNA相关的京都基因与基因组百科全书（KEGG）通路进行了另一项分析。
根据我们的结果，HUCB的扩增导致了miRNA表达水平的改变。本研究为造血细胞二维培养对移植成功的影响提供了信息，以便进行更成功的移植。
