P183 - The effect of blood-derived products on the regenerative potential of adipose-derived stem cells originated from different fat locations
Novel cell sources are currently explored for regenerative joint procedures.
Adipose tissue-derived stem cells (ASCs) have recently been under investigation as a potential cell source.
The aim of this study was to investigate a potentially superior chondrogenic and osteogenic differentiation of ASCs by supplementation with different blood derivatives in in vitro.
Methods and Materials
Fat tissue was harvested from 3 donors, 3 locations each (Hoffa, subcutaneous fat and suprapatellar pouch) during TKA.
ASCs were isolated and cultured in standard growth media.
The expression of specific mesenchymal stem cell markerswasanalysedby flow cytometry (positive: CD90, CD73, CD105).
Platelet-rich plasma (PRP) was prepared either with EDTA (ePRP) or citrate (cPRP) as anticoagulants. HypACT was produced according to the manufacturer´s protocol.
Cells were supplemented with 10% of each blood product in induction medias.
Metabolic activity was assessed by XTT assays.
Differentiation was assessed by staining (osteogenic: alizarin red and dye extraction curve, chondrogenic: Safranin O) and qPCR (osteogenic: ALPL, COL1, RUNX; chondrogenic: SOX9, COL2).
FACS: Cells of all locations expressed MSC markers after isolation (CD90, CD73, CD105).
Metabolic activity: A significant difference was observed in cells supplemented with either cPRP or hypACT in Hoffa cells (fold change day 6: 12 (cPRP) 10 (hypACT) versus 3 (FCS) (p<0,05)).
Osteogenic Differentiation: Alizarin Red Staining and dye extraction curves showed highest mineralization under hypACT and cPRP supplementation both in Hoffa and pouch stem cells (p<0,05).
qPCR did not show significant differences amongst the groups.
Chondrogenic differentiation: qPCR showed highest fold changes amongst Hoffa cells supplemented with ePRP (fold change to day 0: SOX9:38; COL2 1,4).
Preliminary data implypotential beneficial effectsof blood-products on the differentiation potential of ASCs.
Non-anticoagulated serum supplementation may be favourable.
Best differentiation wasobserved withHoffa stem cells.