Prolonged in vitro expansion partially affects phenotypic features and osteogenic potential of ovine amniotic fluid-derived mesenchymal stromal cells |
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| Authors: | Alessia Colosimo Valentina Russo Annunziata Mauro Valentina Curini Marco Marchisio Nicola Bernabò Melissa Alfonsi Mauro Mattioli Barbara Barboni |
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| Affiliation: | 1. Department of Comparative Biomedical Sciences, University of Teramo, Teramo, Italy;2. Department of Biomorphology, University of Chieti, Chieti, Italy;3. Department of Biomedical Sciences, University of Chieti, Chieti, Italy;4. StemTech Group, Chieti, Italy;2. Institut d’Electronique Microelectronique et Nanotechnologie (IEMN UMR Cnrs 8520), University of Lille I, Villeneuve d’Ascq, France;3. Interdisciplinary Research Institute (IRI USR Cnrs 3078), University of Lille I, Villeneuve d’Ascq, France;1. Buck Institute for Age Research, Novato, California, USA;2. Bechman Research Institute, City of Hope, California, USA;3. National Center of Regenerative Medicine, National Institutes of Health, Bethesda, Maryland, USA;1. Department of Hematology, Institute for Internal Medicine; University of Debrecen Medical and Health Science Center, Debrecen, Hungary;2. Institute of Immunology, Rikshospitalet, University of Oslo, Oslo, Norway;1. Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur, Malaysia;5. Department of Obstetrics & Gynecology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur, Malaysia;2. Biotechnology Research Institute, Universiti Malaysia Sabah, Jalan UMS, Kota Kinabalu, Sabah, Malaysia;3. Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, Kuala Lumpur, Malaysia;4. Department of Veterinary Preclinical Sciences, Faculty of Veterinary Medicine, Universiti Putra Malaysia, UPM Serdang, Selangor Darul Ehsan, Malaysia;6. Department of Medical Sciences II, Faculty of Medicine & Health Sciences, Universiti Sains Islam Malaysia, Jalan Pandan Utama, Kuala Lumpur, Malaysia;1. Advanced Centre for Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT, UK;2. Faculty of Science, Engineering and Computing, Kingston University, Penrhyn Road, Kingston upon Thames, Surrey, KT1 2EE, UK;3. London Regenerative Medicine Network, 14a Clerkenwell Green, London, EC1R 0DP, UK;4. Proteus Venture Partners, Portola Valley, CA 94028, USA;5. Centre for Commercialization of Regenerative Medicine, Toronto, ON, M5G 1L5, Canada;6. Harvard Stem Cell Institute, Holyoke Centre, Massachusetts Avenue, Cambridge, MA 02138, USA;1. Department of Orthopaedic Surgery, Ibaraki Prefectural University of Health Sciences, 4669-2 Ami Ami-machi, Inashiki-gun, 300-0394 Ibaraki, Japan;2. Department of Agriculture, Ibaraki University, 3-21-1 Chuo, Ami, 300-0393 Ibaraki, Japan |
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| Abstract: | Background aimsOvine amniotic fluid mesenchymal stromal cells (oAFMSCs) are an emerging alternative source of stem cells to develop pre-clinical cell replacement protocols. For tissue engineering purposes, oAFMSCs can be used either immediately after isolation or after in vitro expansion. However, detailed studies are still required to investigate the advantages and drawbacks of their in vitro expansion.MethodsThe phenotype and osteogenic differentiation potential of oAFMSCs were analyzed in relation to in vitro expansion that was carried out for 20 consecutive passages. Expanded oAFMSCs were analyzed for proliferation index, expression profiles of several surface, pluripotency-associated and HLA antigens, global DNA methylation, telomere length and karyotype. The osteogenic differentiation ability of expanded oAFMSCs was assessed by qualitative and quantitative methods.ResultsExpanded oAFMSCs reduced their proliferative activity after 10 passages and partially modified the expression of surface antigens and the intracellular distribution of pluripotency-associated markers (NANOG, SOX2 and TERT) after 20 passages. The phenotypic alteration of cultured oAFMSCs was associated with a reduction of in vitro osteogenic plasticity. In detail, after 20 passages of cellular expansion, oAFMSCs lost the ability to increase osteocalcin and decreased collagen type I messenger RNA expression. Also, a lower percentage of cells displayed intracellular calcium release after stimulation with salmon calcitonin.ConclusionsThe results presented here suggest that long-term in vitro expansion may cause significant alterations in phenotypic features and plasticity of oAFMSCs, suggesting a careful re-evaluation of in vitro cultural and temporal conditions before employing expanded oAFMSCs for therapeutic purposes. |
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