caa a22 4500 606154965 CHVBK 20210128100601.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s10561-014-9463-8 doi (NATIONALLICENCE)springer-10.1007/s10561-014-9463-8 Defined serum- and xeno-free cryopreservation of mesenchymal stem cells [Elektronische Daten] [Shahla Al-Saqi, Mohammed Saliem, Hernan Quezada, Åsa Ekblad, Aino Jonasson, Outi Hovatta, Cecilia Götherström] Mesenchymal stem cells (MSCs) have vast potential in cell therapy, and are experimentally used in the clinic. Therefore, it is critical to find a serum- and xeno-free cryopreservation method. The aim of this study was to compare two serum- and xeno-free cryoprotectants for MSCs. Adipose tissue MSCs (Ad-MSCs) and bone marrow MSCs (BM-MSCs) were cryopreserved in two cryoprotectants: the defined serum- and xeno-free STEM-CELLBANKER™ (CB) and 10% dimethyl sulfoxide (DMSO) in a xeno-free serum replacement cell culture medium and compared to non-cryopreserved MSCs. MSCs cryopreserved in CB or DMSO had similar morphology and surface marker expression compared to their respective non-cryopreserved MSC. Ad-MSCs and BM-MSC in both cryoprotectant media exhibited reduced mean fluorescence intensity (MFI) for CD105, BM-MSCs for CD73 and Ad-MSC increased MFI for HLA class I compared to non-cryopreserved MSCs. Population doubling time of CB cryopreserved and non-cryopreserved Ad-MSCs was similar (38.1±13.6 and 36.8±12.1h), but somewhat higher when cryopreserved in DMSO (42.2±10.8h). BM-MSCs had higher population doubling time (CB 47.7±11.4 and DMSO 62.3±32.9h respectively, p<0.05) compared to Ad-MSCs. The viability of Ad-MSCs was significantly higher after cryopreservation in CB compared to DMSO (90.4±4.5% vs. 79.9±3.8% respectively). Ad-MSCs and BM-MSCs retained their mesodermal differentiation potential when cryopreserved in both cryoprotectants. The characteristics of Ad-MSCs post-thawing are better preserved by CB than by DMSO in serum- and xeno-free medium. Furthermore, Ad-MSCs and BM-MSCs are differently affected by the cryoprotectants, which may have implications for cell therapy. Springer Science+Business Media Dordrecht, 2014 Mesenchymal stem cells nationallicence Mesenchymal stromal cells nationallicence Adipose nationallicence Bone marrow nationallicence Cryopreservation nationallicence Serum- and xeno-free cryoprotectant nationallicence STEM-CELLBANKER™ nationallicence DMSO nationallicence MSCs : Mesenchymal stem cells nationallicence Ad-MSCs : Adipose tissue-derived MSCs nationallicence CB : STEM-CELLBANKER™ nationallicence DMSO : Dimethyl sulfoxide nationallicence BM-MSCs : Bone marrow-derived MSCs nationallicence HBSS : Hanks balanced salt solution nationallicence PBS : Phosphate-buffered saline nationallicence LDA : Live/dead assay nationallicence PI : Propidium iodide nationallicence PDT : Population doubling time nationallicence DMEM-HG : Dulbecco's modified eagles medium-high glucose nationallicence FBS : Fetal bovine serum nationallicence IBMX : Isobutyl-1-methylxanthine nationallicence Al-Saqi Shahla Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut Saliem Mohammed Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut Quezada Hernan Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, Huddinge, Sweden aut Ekblad Åsa Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut Jonasson Aino Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut Hovatta Outi Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut Götherström Cecilia Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut Cell and Tissue Banking Springer Netherlands 16/2(2015-06-01), 181-193 1389-9333 16:2<181 2015 16 10561 https://doi.org/10.1007/s10561-014-9463-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10561-014-9463-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Al-Saqi Shahla Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut NATIONALLICENCE 700 1- Saliem Mohammed Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut NATIONALLICENCE 700 1- Quezada Hernan Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, Huddinge, Sweden aut NATIONALLICENCE 700 1- Ekblad Åsa Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut NATIONALLICENCE 700 1- Jonasson Aino Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut NATIONALLICENCE 700 1- Hovatta Outi Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut NATIONALLICENCE 700 1- Götherström Cecilia Division of Obstetrics and Gynecology, K57, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, 141 86, Stockholm, Sweden aut NATIONALLICENCE 773 0- Cell and Tissue Banking Springer Netherlands 16/2(2015-06-01), 181-193 1389-9333 16:2<181 2015 16 10561