Cell Viability and Chondrogenic Differentiation Capability of Human Mesenchymal Stem Cells After Iron Labeling with Iron Sucrose

• For evaluation of cell therapy strategies using human mesenchymal stem cells (hMSCs) it is important to be able to trace transplanted cells and their distribution in tissues e.g. cartilage over time. The aim with the study was to determine effects on cell viability, traceability and chondrogenic differentiation of hMSCs after iron labelling with iron sucrose. HMSCs were collected (7 donors, 13-57 years), undergoing spinal surgery. Two sub-sets of experiments were performed. 1)Iron labelling of hMSCs: 1 mg/mL Venofer®(iron sucrose) was added(16 hours) to cultures. hMSCs were examined for uptake of iron sucrose(Preussian blue staining) and cell viability(flow cytometry). 2)Iron labelled hMSCs(passage 4)(n=4, pellet-mass), 200 000 cells/tube were cultured(DMEM-HG) with 10 ng/mL TGFβ and compared to controls(from each donor). The pellets were harvested day 7, 14 and 28. Real time-PCR, IHC and histology were used to evaluate SOX9, ACAN, C6S and COL2A1 expression. Results; mean number of cells containing iron deposits was 98.1 % and mean cell viability 92.7 % (no significant difference compared to unlabelled control cells). Pellets containing iron labelled cells expressed COL2A1 on protein level(all time points), in similar levels as controls and glycosaminoglycan accumulation was observed in iron labelled pellets(day 14 or day 28). Results were supported expression of chondrogenic genes, SOX9, ACAN and COL2A1. The results in vitro indicate that iron sucrose can be used as a cell tracer, for evaluation of cellular distribution in vivo after transplantation of MSCs and thus contribute with important knowledge when exploring new treatment strategies for degenerated cartilaginous tissues.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)

Nyckelord

Mesenchymal stem cells

cell tracer

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