Mesenchymal stromal cells from human umbilical cords display poor chondrogenic potential in scaffold-free three dimensional cultures

Abstract

Many researchers world over are currently investigating the suitability of stromal cells harvested from foetal tissues for allogeneic cell transplantation therapies or for tissue engineering purposes. In this study, we have investigated the chondrogenic potential of mesenchymal stromal cells (MSCs) isolated from whole sections of human umbilical cord or mixed cord (UCSCs-MC), and compared them with cells isolated from synovial membrane (SMSCs), Hoffa's fat pad (HFPSCs) and cartilage. All MSCs were positive for surface markers including CD73, CD90, CD105, CD44, CD146 and CD166, but negative for CD11b, CD19, CD34, CD45 and HLA-DR in addition to CD106 and CD271. Chondrogenic potential of all cell sources was studied using 3D pellet cultures incubated in the presence of different combinations of anabolic substances such as dexamethasone, IGF-1, TGF-β1, TGF-β3, BMP-2 and BMP-7. BMP-2 and dexamethasone in combination with TGF-β1 or TGF-β3 excelled at inducing chondrogenesis on SMSCs, HFPSCs and chondrocytes, as measured by glycosaminoglycans and collagen type II staining of pellets, quantitative glycosaminoglycan expression, quantitative PCR of cartilage signature genes and electron microscopy. In contrast, none of the tested growth factor combinations was sufficient to induce chondrogenesis on UCSCs-MC. Moreover, incubation of UCSCs-MC spheroids in the presence of cartilage pieces or synovial cells in co-cultures did not aid chondrogenic induction. In summary, we show that in comparison with MSCs harvested from adult joint tissues, UCSCs-MC display poor chondrogenic abilities. This observation should alert researchers at the time of considering UCSCs-MC as cartilage forming cells in tissue engineering or repair strategies.

Large-scale secretome analyses unveil the superior immunosuppressive phenotype of umbilical cord stromal cells as compared to other adult mesenchymal stromal cells

“…Macroscopically-normal-looking cartilage, corresponding to cartilage areas looking intact to the eye and presenting no signs of abnormalities, such as tears, scratches, rugose or eroded tissue, was collected from femoral condyles during total knee replacements and was used to isolate human chondrocytes. All cell types were isolated using a mixed enzymatic-explant method, as previously described (Islam et al, 2016;Islam et al, 2017). Briefly, all tissue specimens were washed three times with sterile Dulbecco's phosphate buffered saline (PBS; D8537, Sigma-Aldrich) and minced into small pieces for subsequent enzymatic digestion in collagenase XI solution (≥ 800 units/mg solid; C9407, Sigma-Aldrich) at a final concentration of 1.25 mg/mL at 37 °C on a shaker at 400 rpm.…”

“…In vitro 3D chondrogenic assays of dedifferentiated chondrocytes was performed using a pellet culture method as described previously (Islam et al, 2016). Briefly, chondrocytes were harvested at P3, centrifuged in a conical-bottom 96-well plate for 10 min at 1,100 ×g and incubated in culture medium for 48 h in low oxygen (3 % O 2 ) atmosphere.…”

“…The MS secretomics data (identifier PXD011762) were deposited to the ProteomeXchange Consortium through the PRIDE partner repository (Vizcaíno et al, 2016). All cell types were characterised by MSC surface markers and retained similar characteristics, as shown by Islam et al (2016). The cell secretome established in serum-free CM from each cell type (four unrelated donors per cell type) was analysed by LC-MS/MS proteomics.…”

“…MSC differentiation capacity and immunomodulatory properties are demonstrated in vitro, irrespective of tissue sources (Ghannam et al, 2010). However, in vitro studies show that MSCs from different origins differ in their lineage-specific differentiation capacity and their functional potential (Garcia et al, 2016;Islam et al, 2016;Subramanian et al, 2015). In addition, a systematic review of intra-articular injection of bone marrow MSCs in humans has concluded that articular stem cell therapies are safe (Peeters et al, 2013).…”

Umbilical cord-derived mesenchymal stromal cells: predictive obstetric factors for cell proliferation and chondrogenic differentiation

“…In three-dimensional cultures, cultivated for 3 to 4 weeks in chondrogenic medium supplemented or not with growth factors (such as transforming growth factor (TGF)-β1 and TGF-β3), several groups including our own reported that WJ-MSC showed chondrogenic induction with expression of specific cartilage-related genes and matrix proteins (Sox9, proteoglycans, type 2 collagen, cartilage oligomeric matrix protein (COMP)) [4, 11, 24–26]. However, a recent study was more reserved, showing poor chondrogenesis from MSC isolated from umbilical cord [27]. …”