The effect of medium supplementation and serial passaging on the transcriptome of human adipose-derived stromal cells expanded in vitro

Show simple item record Dessels, Carla Ambele, Melvin Anyasi Pepper, Michael Sean 2020-06-05T06:40:06Z 2020-06-05T06:40:06Z 2019-08
dc.description.abstract BACKGROUND: For adipose-derived stromal cells (ASCs) to be safe for use in the clinical setting, they need to be prepared using good manufacturing practices (GMPs). Fetal bovine serum (FBS), used to expand ASCs in vitro in some human clinical trials, runs the risk of xenoimmunization and zoonotic disease transmission. To ensure that GMP standards are maintained, pooled human platelet lysate (pHPL) has been used as an alternative to FBS. ASCs proliferate more rapidly in pHPL than in FBS, with no significant change in immunophenotype and differentiation capacity. However, not much is known about how pHPL affects the transcriptome of these cells. METHODS: This study investigated the effect of pHPL and FBS on the ASC transcriptome during in vitro serial expansion from passage 0 to passage 5 (P0 to P5). RNA was isolated from ASCs at each passage and hybridized to Affymetrix HuGene 2.0 ST arrays for gene expression analysis. RESULTS: We observed that the transcriptome of ASCs expanded in pHPL (pHPL-ASCs) and FBS (FBS-ASCs) had the greatest change in gene expression at P2. Gene ontology revealed that genes upregulated in pHPL-ASCs were enriched for cell cycle, migration, motility, and cell-cell interaction processes, while those in FBS-ASCs were enriched for immune response processes. ASC transcriptomes were most homogenous from P2 to P5 in FBS and from P3 to P5 in pHPL. FBS- and pHPL-gene-specific signatures were observed, which could be used as markers to identify cells previously grown in either FBS or pHPL for downstream clinical/research applications. The number of genes constituting the FBS-specific effect was 3 times greater than for pHPL, suggesting that pHPL may be a milder supplement for cell expansion. A set of genes were expressed in ASCs at all passages and in both media. This suggests that a unique ASC in vitro transcriptomic profile exists that is independent of the passage number or medium used. CONCLUSIONS: GO classification revealed that pHPL-ASCs are more involved in cell cycle processes and cellular proliferation when compared to FBS-ASCs, which are involved in more specialized or differentiation processes like cardiovascular and vascular development. This makes pHPL a potential superior supplement for expanding ASCs as they retain their proliferative capacity, remain untransformed and pHPL does not affect the genes involved in differentiation in specific developmental processes. en_ZA
dc.description.department Immunology en_ZA
dc.description.department Oral Pathology and Oral Biology en_ZA
dc.description.librarian pm2020 en_ZA
dc.description.sponsorship South African Medical Research Council University Flagship Project, SAMRC Extramural Unit for Stem Cell Research and Therapy and Institute for Cellular and Molecular Medicine of the University of Pretoria. en_ZA
dc.description.uri en_ZA
dc.identifier.citation Dessels, C., Ambele, M.A. & Pepper, M.S. 2019, 'The effect of medium supplementation and serial passaging on the transcriptome of human adipose-derived stromal cells expanded in vitro', Stem Cell Research and therapy, vol. 10, no. 1, art. 253, pp. 1-17. en_ZA
dc.identifier.issn 1757-6512 (online)
dc.identifier.other 10.1186/s13287-019-1370-2
dc.language.iso en en_ZA
dc.publisher BMC en_ZA
dc.rights © 2019 The Author(s). This article is distributed under the terms of the Creative Commons Attribution 4.0 International License. en_ZA
dc.subject Pooled human platelet lysate en_ZA
dc.subject Transcriptome en_ZA
dc.subject Adipose-derived stromal cells (ASCs) en_ZA
dc.subject Good manufacturing practice (GMP) en_ZA
dc.subject Fetal bovine serum (FBS) en_ZA
dc.title The effect of medium supplementation and serial passaging on the transcriptome of human adipose-derived stromal cells expanded in vitro en_ZA
dc.type Article en_ZA

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