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Microarray profiling of in vitro expanded FRCs cultured alone or with activated T cells

ABSTRACT: Microarray profiling of in vitro expanded FRCs cultured alone or with activated T cells

PROVIDER: PRJNA564111 | ENA |

REPOSITORIES: ENA

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Microarray profiling of in vitro expanded FRCs cultured alone or with activated T cells

Project description:To investigate whether FRCs express molecules capable of promoting the functions of activated T cells, we expanded FRCs from primary lymph node stromal cell (LNSC) cultures as previously described (Lukacs-Kornek et al., Nature Immunology, 2011), and then cultured them alone or with splenocytes activated with soluble antibody (0.25μg/ml) against CD3 (anti-CD3) and anti-CD28 for 16 hours. FRCs co-cultured with activated T cells upregulated expression of genes encoding molecules known to dampen T cell function such as Arg1, CD274 and Nos2. However, in response to activated T cells, FRCs also upregulated molecules with immunostimulatory capabilities such as Icosl, Cd40 and Il6.

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RNA seq dataset of parental and reserve cells (FRCs Cycle 1 and SRCs Cycle 3)

Project description:Loss of regenerative capacity during ex vivo expansion of muscle stem cells hampers the development of cell-based therapies for skeletal muscle disorders. Here, we developed a method to isolate regenerative reserve cell fractions from expanded murine primary cultures using differential adhesion properties and the induction of subsequent cycles of proliferation and differentiation. Fast-adhering reserve cells (FRCs) were enriched for cells expressing myogenin and contributed efficiently to muscle regeneration following transplantation in immunodeficient hosts. Slow adhering reserve cells (SRCs) generated higher number of PAX7+ cells under differentiating conditions and predominantly regenerated muscle after a secondary injury, suggesting engraftment as muscle stem cells. Transcriptomic analysis revealed that FRCs were enriched for markers of myogenic differentiation, while SRCs were characterized by a distinctive set of cell-adhesion genes. Overall, reserve cells provide a valuable source for the study and identification of molecular determinants of regeneration using ex vivo-expanded muscle cells.

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