Differential ability of MSCs isolated from placenta and cord as feeders for supporting ex vivo expansion of umbilical cord blood derived CD34 + cells

Stem Cell Research & Therapy

Lalita Limaye¹  Vaijayanti Kale¹  Darshana Kadekar¹ 

[1]Stem Cell Laboratory, National Centre for Cell Science, University of Pune Campus, Ganeshkhind, Pune 411007, Maharashtra, India

关键词: Placental mesenchymal stem cells;    Cord mesenchymal stem cells;    Ex vivo HSC expansion;   

Others  :  1234585
DOI  :  10.1186/s13287-015-0194-y

 received in 2015-03-03, accepted in 2015-09-28,  发布年份 2015

【 摘 要 】

Introduction

Ex vivo expansion of umbilical cord blood (UCB) is attempted to increase cell numbers to overcome the limitation of cell dose. Presently, suspension cultures or feeder mediated co-cultures are performed for expansion of hematopoietic stem cells (HSCs). Mesenchymal stem cells (MSCs) have proved to be efficient feeders for the maintenance of HSCs. Here, we have established MSCs-HSCs co-culture system with MSCs isolated from less invasive and ethically acceptable sources like umbilical cord tissue (C-MSCs) and placenta (P-MSCs). MSCs derived from these tissues are often compared with bone marrow derived MSCs (BM-MSCs) which are considered as a gold standard. However, so far none of the studies have directly compared C-MSCs with P-MSCs as feeders for ex vivo expansion of HSCs. Thus, we for the first time performed a systematic comparison of hematopoietic supportive capability of C and P-MSCs using paired samples.

Methods

UCB-derived CD34 ⁺cells were isolated and co-cultured on irradiated C and P-MSCs for 10 days. C-MSCs and P-MSCs were isolated from the same donor. The cultures comprised of serum-free medium supplemented with 25 ng/ml each of SCF, TPO, Flt-3 L and IL-6. After 10 days cells were collected and analyzed for phenotype and functionality.

Results

C-MSCs and P-MSCs were found to be morphologically and phenotypically similar but exhibited differential ability to support ex vivo hematopoiesis. Cells expanded on P-MSCs showed higher percentage of primitive cells (CD34 ⁺ CD38 ⁻ ), CFU (Colony forming unit) content and LTC-IC (Long term culture initiating cells) ability. CD34 ⁺cells expanded on P-MSCs also exhibited better in vitro adhesion to fibronectin and migration towards SDF-1α and enhanced NOD/SCID repopulation ability, as compared to those grown on C-MSCs. P-MSCs were found to be closer to BM-MSCs in their ability to expand HSCs. P-MSCs supported expansion of functionally superior HSCs by virtue of reduction in apoptosis of primitive HSCs, higher Wnt and Notch activity, HGF secretion and cell-cell contact. On the other hand, C-MSCs facilitated expansion of progenitors (CD34 ⁺ CD38 ⁺ ) and differentiated (CD34 ⁻ CD38 ⁺ ) cells by secretion of IL1-α, β, MCP-2, 3 and MIP-3α.

Conclusions

P-MSCs were found to be better feeders for ex vivo maintenance of primitive HSCs with higher engraftment potential than the cells expanded with C-MSCs as feeders.

【 授权许可】

   
2015 Kadekar et al.

【 图 表 】

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