Department of Neurology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA.
Multiple Sclerosis Center of Excellence, East VA Maryland Health Care System, Baltimore, MD, 21201, USA.
Stem Cell Res Ther. 2018 May 2;9(1):127. doi: 10.1186/s13287-018-0858-5.
Adult bone marrow contains stem cells that replenish the myeloid and lymphoid lineages. A subset of human and mouse CD34 bone marrow stem cells can be propagated in culture to autonomously express embryonic stem cell genes and embryonic germ layer lineage genes. The current study was undertaken to determine whether these CD34 stem cells could be obtained from human blood, whether gene expression could be modulated by culture conditions and whether the cells produce insulin.
Human peripheral blood buffy coat cells and mobilized CD34 cells from human blood and from blood from C57Bl/6 J mice were cultured in hybridoma medium or neural stem cell induction medium supplemented with interleukin (IL)-3, IL-6, and stem cell factor (SCF). Changes in mRNA and protein expression were assessed by Western blot analysis and by immunohistochemistry. Mass spectrometry was used to assess insulin production.
We were able to culture CD34 cells expressing embryonic stem cell and embryonic germ layer lineage genes from adult human peripheral blood after standard mobilization procedures and from mouse peripheral blood. Gene expression could be modulated by culture conditions, and the cells produced insulin in culture.
These results suggest a practical method for obtaining large numbers of CD34 cells from humans to allow studies on their potential to differentiate into other cell types.
成人骨髓中含有可补充髓系和淋巴系的干细胞。人类和小鼠 CD34 骨髓干细胞的一个亚群可在培养中增殖，自主表达胚胎干细胞基因和胚胎生殖层谱系基因。本研究旨在确定这些 CD34 干细胞是否可从人血液中获得，培养条件是否可调节基因表达，以及细胞是否产生胰岛素。
用人外周血白细胞层细胞和人外周血及 C57Bl/6J 小鼠外周血中的动员 CD34 细胞，在杂交瘤培养基或神经干细胞诱导培养基中培养，补充白细胞介素 (IL)-3、IL-6 和干细胞因子 (SCF)。通过 Western blot 分析和免疫组织化学评估 mRNA 和蛋白质表达的变化。采用质谱法评估胰岛素的产生。
我们能够在标准动员程序后，从成人外周血和小鼠外周血中培养表达胚胎干细胞和胚胎生殖层谱系基因的 CD34 细胞。基因表达可通过培养条件调节，细胞在培养中产生胰岛素。
这些结果表明，从人类获得大量 CD34 细胞的一种实用方法，可用于研究其分化为其他细胞类型的潜力。
