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Comparative analysis of multilineage properties of mesenchymal stromal cells derived from fetal sources shows an advantage of mesenchymal stromal cells isolated from cord blood in chondrogenic differentiation potential
Institution:1. Dulbecco Telethon Institute at Tettamanti Research Center, Pediatric Department, University of Milano-Bicocca, Monza, Italy;2. Tettamanti Research Center, Pediatric Department, University of Milano-Bicocca, San Gerardo Hospital, Monza, Italy;3. Department of Obstetrics and Gynecology, University of Milano-Bicocca, Monza, Italy;4. Department of Molecular Medicine, Sapienza University, Rome, Italy;5. Department of Oncology, Flow Cytometry Unit, IRCCS Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy;6. Pediatric Department, University of Milano-Bicocca, Fondazione MBBM/San Gerardo Hospital, Monza, Italy
Abstract:Background aimsCord blood (CB) and amniotic fluid (AF) could represent new and attractive mesenchymal stromal cell (MSC) sources, but their potential therapeutic applications are still limited by lack of standardized protocols for isolation and differentiation. In particular, chondrogenic differentiation has never been deeply investigated.MethodsMSCs were obtained from CB and AF samples collected during cesarean sections at term and compared for their biological and differentiation properties, with particular interest in cartilage differentiation, in which quantitative real-time polymerase chain reaction and immunohistochemical analyses were performed to evaluate the expression of type 2 collagen, type 10 collagen, SRY-box9 and aggrecan.ResultsWe were able to isolate MSCs from 12 of 30 (40%) and 5 of 20 (25%) CB and AF units, respectively. Fluorescence in situ hybridization analysis indicated the fetal origin of isolated MSC strains. Both populations expressed mesenchymal but not endothelial and hematopoietic markers, even though we observed a lower expression of human leukocyte antigen (HLA) I in CB-MSCs. No differences in proliferation rate and cell cycle analysis could be detected. After osteogenic induction, both populations showed matrix mineralization and typical marker expression. Under chondrogenic conditions, pellets derived from CB-MSCs, in contrast with AF-MSCs pellets, were significantly larger, showed cartilage-like morphology and resulted positive for chondrocyte-associated markers, such as type 2 collagen, type 10 collagen, SRY-box9 and aggrecan.ConclusionsOur results show that CB-MSCs and AF-MSCs collected at term differ from each other in their biological and differentiation properties. In particular, only CB-MSCs showed a clear chondrogenic potential and thus could represent an ideal candidate for cartilage-tissue engineering.
Keywords:amniotic fluid  chondrogenic differentiation  cord blood  mesenchymal stromal cells
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