Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8464128 | Cryobiology | 2018 | 33 Pages |
Abstract
The use of fetal bovine serum (FBS) as a cryopreservation supplement is not suitable for the banking of mesenchymal stem cells (MSCs) due to the risk of transmission of disease as well as xenogeneic immune reactions in the transplanted host. Here, we investigated if human serum albumin (HSA), human serum (HS), or knockout serum replacement (KSR) can replace FBS for the cryopreservation of MSCs. In addition, we examined the characteristics of MSCs after multiple rounds of cryopreservation. Human adipose-derived stem cells (ASCs) cryopreserved with three FBS replacements, 9% HSA, 90% HS, or 90% KSR, in combination with 10% dimethyl sulfoxide (Me2SO) maintained stem cell properties including growth, immunophenotypes, gene expression patterns, and the potential to differentiate into adipogenic, osteogenic, and chondrogenic lineages, similar to ASCs frozen with FBS. Moreover, the immunophenotype, gene expression, and differentiation capabilities of ASCs were not altered by up to four freeze-thaw cycles. However, the performance of three or four freeze-thaw cycles significantly reduced the proliferation ability of ASCs, as indicated by the longer population doubling time and reduced colony-forming unit-fibroblast frequency. Together, our results suggest that HSA, HS, or KSR can replace FBS for the cryopreservation of ASCs, without altering their stemness, and should be processed with no more than two freeze-thaw cycles for clinical approaches.
Keywords
FACSComplement Factor DBGNMe2SOKSRCOMPBiglycanOCNPPARγMSCRT-PCRRUNX2CFU-FGAPDHDPBSHSAASCHSCDCNFITCFBSC/EBPαCCAAT/enhancer-binding protein alphahuman serum albuminOsteocalcinClinical useknockout serum replacementfluorescence-activated cell sortingdecorinCFDDimethyl sulfoxideHuman serumfetal bovine serumCryopreservationHematopoietic stem cellAdipose-derived stem cellhuman adipose-derived stem cellsMesenchymal stem cellRunt-related transcription factor 2phycoerythrinDulbecco's phosphate-buffered salinefluorescein isothiocyanatereverse transcription polymerase chain reactionCartilage oligomeric matrix proteinPropidium iodideFreeze-thaw cyclesPeroxisome proliferator-activated receptor gammaglyceraldehyde-3-phosphate dehydrogenase
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Authors
Seah Park, Dong Ryul Lee, Ji Sun Nam, Chul Woo Ahn, Haekwon Kim,