Du Zheng, Wang Ziyan, Zhang Weiwei, Cai Haibo, Tan Wen-Song
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
Cell Prolif. 2015 Jun;48(3):293-300. doi: 10.1111/cpr.12186. Epub 2015 Apr 21.
Stem cell factor (SCF) is essential in the haematopoietic stem cells (HSCs) niche, and is therefore used extensively in haematopoietic stem and progenitor cells (HSPCs) ex vivo expansion. However, in the literature, dose and schedule of SCF feeding varies widely. We previously proposed a novel SCF feeding regimen with proven effectiveness for HSPCs expansion; however, physiological function of expanded cells with this SCF feeding regimen required further research.
CD34(+) cells were cultured with or without SCF supplementation in serum-free medium for 10 days. Expanded cells were transplanted into sublethally irradiated non-obese diabetic/severe combined immune-deficient (NOD/SCID) mice. Engraftment and multilineage reconstitution of transplanted cells were determined. Also, clonogenic potential of engrafted cells was analysed.
Cells, both cultured with and without SCF supplementation, successfully engrafted and reconstituted blood cell lineages in NOD/SCID mice. However, level of engraftment and multilineage reconstitution reduced when cells were expanded without SCF supplementation. Meanwhile, frequencies of colony-forming cells (CFCs) amongst bone marrow cells were higher in mice transplanted with CD34(+) cells expanded with SCF supplementation.
Reconstitution capacity reduced when CD34(+) cells were expanded without SCF supplementation, though this feeding regimen did not have any effect on cell expansion. This finding suggested that SCF was essential for preserving NOD/SCID reconstitution capacity of ex vivo expanded CD34(+) cells.
干细胞因子(SCF)在造血干细胞(HSC)微环境中至关重要,因此广泛用于造血干祖细胞(HSPC)的体外扩增。然而,在文献中,SCF添加的剂量和方案差异很大。我们之前提出了一种新型的SCF添加方案,已证实其对HSPC扩增有效;然而,采用这种SCF添加方案扩增的细胞的生理功能需要进一步研究。
将CD34(+)细胞在无血清培养基中培养10天,添加或不添加SCF。将扩增后的细胞移植到经亚致死剂量照射的非肥胖糖尿病/重症联合免疫缺陷(NOD/SCID)小鼠体内。测定移植细胞的植入和多谱系重建情况。此外,分析植入细胞的克隆形成潜力。
添加和未添加SCF培养的细胞均成功植入NOD/SCID小鼠体内并重建血细胞谱系。然而,未添加SCF扩增的细胞,其植入水平和多谱系重建能力降低。同时,在移植了添加SCF扩增的CD34(+)细胞的小鼠中,骨髓细胞中集落形成细胞(CFC)的频率更高。
未添加SCF扩增CD34(+)细胞时,重建能力降低,尽管这种添加方案对细胞扩增没有任何影响。这一发现表明,SCF对于维持体外扩增的CD34(+)细胞的NOD/SCID重建能力至关重要。