Unknown,Transcriptomics,Genomics,Proteomics

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Expression data from mesenchymal stromal cells isolated from the umbilical cord tissue (UCXM-BM-.) and cultivated in ATMP-compatible media

ABSTRACT: Standardization of MSC manufacturing is urgently needed to facilitate comparison of clinical trial results. Here, we compare gene expression of MSC generated by the adaptation of a proprietary method for isolation and cultivation of a specific umbilical cord tissue-derived population of Mesenchymal Stromal Cells (MSCs) The adaptation focused on different stages of production, from cell isolation steps to cell culturing and preservation. The origin and quality of reagents were considered and steps for avoiding microbiological and endotoxin contamination of the final cell product were implemented. Cell isolation efficiency, MSC surface markers and genetic profiles originating from the use of different medium supplements were compared. The ATMP-compliant UCXM-BM-. product was also cryopreserved avoiding the use of DMSO, an added benefit in the use of these cells as an ATMP. Cells were analysed for expansion capacity and longevity. The final cell product was further characterised by flow cytometry, differentiation potential, and tested for contaminants at various passages. Gene expression profiles, genetic stability and immune properties were analysed. human umbilical cord tissue-derived MSCs (UCXM-BM-.) were isolated using clinical grade enzymes and cultivated in flask or bags using M-NM-1-MEM basal medium with 1 g/L glucose and 2 mM glutamine supplemented with either FBS,human serum or human platelet lysate.

ORGANISM(S): Homo sapiens

SUBMITTER: Robert Geffers

PROVIDER: E-GEOD-51869 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:We have previously reported that the deficiency of p53 alone or in combination with Rb (Rb-/- p53-/-) in adipose-derived MSCs (ASCs) promotes leiomyosarcoma-like tumors in vivo. Here, we hypothesized that the source of MSCs and/or the cell differentiation stage could determine the phenotype of sarcoma development. To investigate whether there is a link between the source of MSCs and sarcoma phenotype, we generated p53-/- and Rb-/-p53-/- MSCs from bone marrow (BM-MSCs). Both genotypes of BM-MSCs initiated leiomyosarcoma formation similar to p53-/- and Rb-/-p53-/- ASCs. In addition, gene expression profiling revealed a link between p53- or Rb-p53-deficient BM-MSCs and ASCs and muscle-associated sarcomagenesis. These data suggest that the tissue source of MSC does not seem a crucial factor in the development of a particular sarcoma phenotype. To analyze whether the differentiation stage defines the sarcoma phenotype, BM-MSCs and ASCs were induced to differentiate towards the osteogenic lineage, and both p53 and Rb were excised using Cre-expressing adenovectors at different stages along osteogenic differentiation. Regardless of the level of osteogenic commitment, the inactivation of Rb and p53 in BM-MSC-derived, but not in ASC-derived, osteogenic progenitors gave rise to osteosarcoma-like tumors which could be serially transplanted. This indicates that the osteogenic differentiation stage of BM-MSCs imposes the phenotype of in vivo sarcoma development, and that BM-MSC-derived osteogenic progenitors rather than undifferentiated BM-MSCs, undifferentiated ASCs or ASC-derived osteogenic progenitors, represent the cell of origin for osteosarcoma development. To analyse whether the BM-MSC and Fat-MSC (ASC) differentiation stage may define the sarcoma phenotype, RbloxP/loxPp53loxP/loxP BM-MSCs and ASCs were induced to differentiate towards the osteogenic lineage and both Rb and p53 were excised with adenoviral vectors expressing the Cre-recombinase gene (Ad-CMV-Cre) at different stages (day 0 and 10) along osteogenic differentiation. NSG mice were inoculated subcutaneously with 5M-CM-^W10^6 mutant cells. Animals were killed when tumors reached 1 cm3 or 150 days after infusion. Some of the obtained tumors were mechanically disaggregated to establish ex vivo MSC-transformed cell lines. Gene expression analysis was performed using: WT BM-MSCs and ASCs, Rb-/-p53-/- BM-MSCs and ASCs previously differentiated to the osteogenic lineage for 10 days and a tumor cell line derived from p53-/-Rb-/- BM-MSC differentiated to the osteogenic lineage for 10 days.

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Expression data from mesenchymal stromal cells isolated from the umbilical cord tissue (UCXM-BM-.) and cultivated in ATMP-compatible media

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