Project description:Spermatogonial stem cells (SSCs) undergo self-renewal division to maintain spermatogenesis. In this study, we used SSCs for long-term cryopreservation in space flight. We derived germline stem cell (GS) cultures from DBA/2 mouse testes (Kanatsu-Shinohara et al., 2003, Biol. Reprod. 69, 216-616). GS cells were cryopreserved in the ISS for 6 months. Frozen GS cells were thawed upon return to Earth and tranplanted into the seminifelous tubules of infertile W mice. Recipients were housed with wild-type C57BL/6 females, and livers of F1 offspring were analyzed for gene expression.
Project description:Spermatogonial stem cells (SSCs) undergo self-renewal division to maintain spermatogenesis. In this study, we used SSCs for long-term cryopreservation in space flight. We derived germline stem cell (GS) cultures from DBA/2 mouse testes by supplementing GDNF and FGF2 (Kanatsu-Shinohara et al., 2003, Biol. Reprod. 69, 216-616). GS cells were cryopreserved in the ISS for 6 months. Frozen GS cells were thawed upon return to Earth and their gene expression profiles were analysed.
Project description:Spermatogonial stem cells (SSCs) undergo self-renewal division to maintain spermatogenesis. In this study, we used SSCs for long-term cryopreservation in space flight. We derived germline stem cell (GS) cultures from DBA/2 mouse testes by supplementing GDNF and FGF2 (Kanatsu-Shinohara et al., 2003, Biol. Reprod. 69, 216-616). GS cells were cryopreserved in the ISS for 6 months. Frozen GS cells were thawed upon return to Earth and their gene expression profiles were analysed.
Project description:The in vitro derivation and propagation of spermatogonial stem cells (SSCs) from pluripotent stem cells (PSCs) is a key goal in reproductive science. We show here that when aggregated with embryonic testicular somatic cells (reconstituted testes), primordial germ cell-like cells (PGCLCs) induced from mouse embryonic stem cells differentiate into spermatogonia-like cells in vitro and are expandable as cells that resemble germline stem cells (GSCs), a primary cell line with SSC activity. Remarkably, GSC-like cells (GSCLCs), but not PGCLCs, colonize adult testes and, albeit less effectively than GSCs, contribute to spermatogenesis and fertile offspring. Whole-genome analyses reveal that GSCLCs exhibit aberrant methylation at vulnerable regulatory elements, including those critical for spermatogenesis, which may restrain their spermatogenic potential. Our study establishes a strategy for the in vitro derivation of SSC activity from PSCs, which, we propose, relies on faithful epigenomic regulation.
Project description:The in vitro derivation and propagation of spermatogonial stem cells (SSCs) from pluripotent stem cells (PSCs) is a key goal in reproductive science. We show here that when aggregated with embryonic testicular somatic cells (reconstituted testes), primordial germ cell-like cells (PGCLCs) induced from mouse embryonic stem cells differentiate into spermatogonia-like cells in vitro and are expandable as cells that resemble germline stem cells (GSCs), a primary cell line with SSC activity. Remarkably, GSC-like cells (GSCLCs), but not PGCLCs, colonize adult testes and, albeit less effectively than GSCs, contribute to spermatogenesis and fertile offspring. Whole-genome analyses reveal that GSCLCs exhibit aberrant methylation at vulnerable regulatory elements, including those critical for spermatogenesis, which may restrain their spermatogenic potential. Our study establishes a strategy for the in vitro derivation of SSC activity from PSCs, which, we propose, relies on faithful epigenomic regulation.
2016-12-06 | GSE76245 | GEO
Project description:HIV-1 RNA sequencing from PBMCs cryopreserved for more than twenty years
Project description:Multipotent spermatogonial stem cells (mSSCs) derived from SSCs are a potential new source of individualized pluripotent cells in regenerate medicine such as ESCs. We hypothesized that the culture-induced reprogramming of SSCs was mediated by a mechanism different from that of iPS, and was due to up-regulation of specific pluripotency-related genes during cultivation. Through a comparative analysis of expression profile data, we try to find cell reprogramming candidate factors from mouse spermatogonial stem cells. We used microarrays to analyze the gene expression profiles of culture-induced reprogramming converting unipotent spermatogonial stem cells to pluripotent spermatogonial stem cells. Three types of spermatogonial stem cells were mechanically collected according to morphological criteria for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Mouse spermatogonial stem cells (SSCs) continuously self-renew on the feeder layers in serum-free culture medium supplemented with glial cell line-derived neurotrophic factor and fibroblast growth factor 2. To identify novel nuclear proteins involved in SSC maintenance, comparative proteomic profiling of nuclear proteins was performed between self-renewing and differentiation-initiated SSCs in culture. The self-renewing SSC cultures were established from C57BL/6 mouse testes. Nuclear fractions from self-renewing SSC cultures treated with ethanol as a vehicle control (spermatogonial stem cells) and differentiation-initiated SSC cultures treated with 0.3 μM retinoic acid for 24 h (spermatogonial progenitor cells) were isolated for proteomic analysis.
Project description:In vitro and in vivo aging of mouse spermatogonial stem cells alters stem cell function based on quantitative spermatogonial stem cell transplantation analyses. We used microarrays to identify differential gene expression in vitro and in vivo aged spermatogonial stem cells to identify potential causes of observed phenotypic differences in aged spermatogonial stem cell function. Spermatogonial stem cells were isolated from young and serial-transplanted aged mouse donors and cultured for short and long periods. Spermatogonial stem cells were isolated from cultures and subjected to microarray analysis to identify differential gene expression.
