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

Abstract

IntroductionEx 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.MethodsUCB-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.ResultsC-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.ConclusionsP-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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0194-y) contains supplementary material, which is available to authorized users.

Conditioned Medium from Placental Mesenchymal Stem Cells Reduces Oxidative Stress during the Cryopreservation of Ex Vivo Expanded Umbilical Cord Blood Cells

“…According to our earlier findings [22], we observed that P-MSCs-CM had superior cryoprotective activity than C-MSCs-CM. This difference may be attributed to the distinct secretory profiles exhibited by C and P-MSCs as described in this report [22].…”

“…This difference may be attributed to the distinct secretory profiles exhibited by C and P-MSCs as described in this report [22]. Although, P and C-MSCs are in close anatomical proximity, both exhibited differences in their characteristics, such as higher concentration of HGF in P-MSCs- CM compared to C-MSCs-CM [22]. HGF is known to have anti-oxidant effect [20,21] which might have contributed to the superior cryoprotection provided by the P-MSCs-CM.…”

“…MSCs were isolated from cord (n = 7) and placenta (n = 7) as described in reference [22]. Briefly, P-MSCs were isolated specifically from chorionic and decidual part of the placenta after removal of amniotic membrane and C-MSCs were obtained after removal of vitelline arteries and vein from Wharton’s jelly and the mesenchyme present within the different layers of cord which we isolate by squeezing the tissue mechanically.…”

Multiplacenta derived stem cell/cytokine treatment increases survival time in a mouse model with radiation-induced bone marrow damage

“…Thus, we transplanted the stem cells combined with placentapooled cytokines. Placenta-derived mesenchymal and hematopoietic stem cells as well as cytokines are homologous (Kadekar et al 2015), and the cytokines combined with DMEM/F-12 can maintain the survival of the mesenchymal and hematopoietic stem cells.…”

Inhibin α‐subunit inhibits BMP9‐induced osteogenic differentiation through blocking BMP/Smad signal and activating NF‐κB signal in mesenchymal stem cells

“…Marrow stromal cells, firstly being identified in bone, can differentiate into multiple lineages, such as osteoblasts, adipocyte, chondrocytes, and myoblasts in vitro . With the stemness of self‐renewal and multipotential differentiation ability, MSCs from other various tissues are more and more recognized as seed cells for bone remodeling . Due to the features of expansion and multiple differentiation potential in vitro, MSCs are supposed to be a core of the tissue bone engineering …”