Comparison of cord blood CD34+stem cell expansion in coculture with mesenchymal stem cells overexpressing SDF-1 and soluble/membrane isoforms of SCF
Comparison of cord blood CD34+stem cell expansion in coculture with mesenchymal stem cells overexpressing SDF-1 and soluble/membrane isoforms of SCF
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DOI：
10.1002/jcb.28797
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发表时间：
2019-09-01
影响因子：
4
通讯作者：
Atashi, Amir
中科院分区：
文献类型：
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作者：
Ajami, Mansoureh;Soleimani, Masoud;Atashi, Amir
目的脐带血造血干细胞(UCB HSCs)因其无创性采集、扩增能力强、移植耐受能力强等优点，被认为可用于治疗恶性血液病。另一方面，这些细胞最大的局限性是它们的CD34+造血干细胞数量不足，导致植入延迟。本研究的目的是通过与高表达基质细胞衍生因子-1、可溶性和膜型干细胞因子(sSCF/mscf)的间充质干细胞(MSCs)共培养扩增CD34+HSCs。MSCs能够维持高表达的细胞因子的结构和功能，有利于CD34+HSC的扩增。方法从脐血单个核细胞中分离出MSCs和CD34+HSCs。将sSCF、mscf和SDF-1表达载体中的一个或多个与脐血间充质干细胞进行核重组。然后将分离的CD34+HSCs与10组携带核的MSCs在含有TPO和Flt3L的培养液中加或不加SCF(3F组或2F组)共培养。经修饰的MSCs与CD34+HSCs共培养后，检测CD34+HSCs的数量、克隆能力以及著名的HSCs调控和干性基因(CXCR4、HOXB4、BMI1和SALL4)的转录水平。结果3F组CD34+HSCs在高表达mscf/sSCF/SDF-1的MSCs上扩增(4.73+/-0.26倍)、克隆形成能力(5.3+/-0.25倍)、CXCR4、HOXB4和BMI1转录水平(3.49+/-0.13倍、9.49+/-0.78倍和11.6+/-0.9倍)均显著增加(P<0.05)。结论脐血间充质干细胞在共培养体系中过表达SCF和SDF-1，对脐血间充质干细胞的扩增有明显的促进作用。此外，在HSC扩增培养液中，过量表达sSCF/mscf或mscf/sSCF/SDF-1的核修饰的MSC可以替代rhu SCF。
Aim Umbilical cord blood hematopoietic stem cells (UCB HSCs) have been considered for the treatment of hematological malignancies due to their noninvasive collection, greater capacity of expansion, and remarkable tolerance for HLA mismatch in transplantation. On the other hand, the most considerable limitation of these cells is their inadequate amount of CD34 + HSCs which leads to delayed engraftment. The aim of this study was the expansion of CD34 + HSCs by coculturing with mesenchymal stem cells (MSCs) overexpressing stromal cell-derived factor-1, soluble and membrane isoforms of stem cell factor (sSCF/mSCF). Keeping structure and function of overexpressed cytokines by MSCs was expected which could beneficially affect the CD34 + HSC expansion. Methods MSCs and CD34 + HSCs were isolated from UCB mononuclear cells. UCB MSCs were nucleofected with one or more of sSCF, mSCF, and SDF-1 expression vectors. Isolated CD34 + HSCs were then cocultured with nucleofected MSCs in 10 groups in culture medium containing TPO and Flt3L with or without SCF (3F or 2F groups). Then the CD34 + HSCs numeration, clonogenic capacity, and transcriptional levels of well-known HSCs regulatory and stemness genes (CXCR4, HOXB4, BMI1, and SALL4) were assessed following coculture with modified MSCs. Results CD34 + HSCs which expanded on MSCs overexpressing mSCF/sSCF/SDF-1 in the 3F group showed the most significant increase in the expansion (4.73 +/- 0.26 fold), clonogenic capacity (5.3 +/- 0.25 fold) and also transcriptional levels of CXCR4, HOXB4, and BMI1 (3.49 +/- 0.13, 9.49 +/- 0.78, and 11.6 +/- 0.9 fold), respectively (P < 0.05). Conclusion Overexpression of SCF and SDF-1 by UCB MSCs in coculture system has efficient effect on UCB HSCs expansion. Furthermore nucleofected MSC overexpressing either sSCF/mSCF or mSCF/ sSCF /SDF-1 could substitute the rhu SCF in HSC expansion culture medium.