Fertile offspring derived from mouse spermatogonial stem cells cryopreserved for more than 14 years
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE35056 | GEO | 2012/01/13
SECONDARY ACCESSION(S): PRJNA150839
REPOSITORIES: GEO
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Project description:To evaluate the possible deleterious affects prolonged cryopreservation and assisted reproductive technologies may have on the DNA of F0 progeny, we used array-based comparative genomic hybridization (CGH) for analysis of genomic DNA. ICSI-derived progeny were generated from ♂ (ZFLacZ sperm) x ♀ (DBA x C57BL/6 oocyte), where the ZFLazZ transgenic mice were originally generated from an SJL x C57BL/6 background.
2012-01-13 | E-GEOD-35056 | biostudies-arrayexpress
Project description:BackgroundApproximately 80% of childhood cancers can now be cured but a side effect of treatment results in about one-third of the surviving boys being infertile or severely subfertile when they reach reproductive age. Currently, more than 1 in 5000 men of reproductive age who are childhood cancer survivors suffer from this serious quality of life problem. It is possible to obtain a testicular biopsy before treatment to preserve the spermatogonial stem cells (SSCs) of the male by cryopreservation, but the results of long-term storage of SSCs on their subsequent functional ability to generate normal offspring has not been examined in any mammalian species. Moreover, it will be necessary to increase the number of these cryopreserved SSCs to remove any contaminating malignant cells and assure regeneration of spermatogenesis.Methods and resultsIn this report, we demonstrate that long-term cryopreservation (>14 years) of testis cells from mouse, rat, rabbit and baboon safeguards SSC viability, and that these cells can colonize the seminiferous tubules of recipient testes. Moreover, mouse and rat SSCs can be cultured and re-establish complete spermatogenesis, and fertile mouse progeny without apparent genetic or epigenetic errors were generated by the sperm produced.ConclusionsThese findings provide a platform for fertility preservation in prepubertal boys undergoing gonadotoxic treatments.
| S-EPMC3329194 | biostudies-literature
Project description:Treatment of cancer in children is increasingly successful but leaves many prepubertal boys suffering from infertility or subfertility later in life. A current strategy to preserve fertility in these boys is to cryopreserve a testicular biopsy prior to treatment with the expectation of future technologies allowing for the reintroduction of stem cells and restoration of spermatogenesis. Spermatogonial stem cells form the basis of male reproduction, differentiating into all germ cell types, including mature spermatozoa and can regenerate spermatogenesis following transplantation into an infertile testis. Here we demonstrate for the first time that rat spermatogonial stem cells frozen for more than 20 years can be transplanted into recipient mice and produce all differentiating germ cell types. However, compared with freshly-isolated cells or those frozen for a short period of time, long frozen cells do not colonize efficiently and showed reduced production of spermatids. Single cell RNA sequencing reveled similar profiles of gene expression changes between short and long frozen cells as compared with fresh immediately after thawing. Conversely, following transplantation, long frozen samples showed enhanced stem cell signaling in the undifferentiated spermatogonia compartment, consistent with self-renewal and a lack of differentiation. In addition, long frozen samples showed fewer round spermatids with detectable protamine expression, suggesting a partial block of spermatogenesis after meiosis resulting in a lack of elongating spermatids. These findings strongly suggest that prolonged cryopreservation can impact the success of transplantation to produce spermatogenesis, which may not be revealed by analysis of the cells immediately after thawing.
2022-03-15 | GSE182438 | GEO
Project description:This study investigated which genes regarding root resorption are upregulated by cryopreservation and whether cryopreservation affects the expression of Macrophage –colony stimulating factor. We manufactured the customized template which was made of genes selected regarding root resorption including osteoprotegerin (OPG), receptor activator of nuclear factor-kappa B ligand (RANKL), RANKL’s cognate receptor (RANK), macrophage colony-stimulating factor (M-CSF), interleukin-1β (IL-1β), and tumor necrosis factor-alpha (TNF-α), and bone morphogenetic proteins (BMP) and analyzed gene expression.
2013-01-19 | GSE43611 | GEO
Project description:This study investigated which genes regarding root resorption are upregulated by cryopreservation and whether cryopreservation affects the expression of Macrophage M-bM-^@M-^Scolony stimulating factor. We manufactured the customized template which was made of genes selected regarding root resorption including osteoprotegerin (OPG), receptor activator of nuclear factor-kappa B ligand (RANKL), RANKLM-bM-^@M-^Ys cognate receptor (RANK), macrophage colony-stimulating factor (M-CSF), interleukin-1M-NM-2 (IL-1M-NM-2), and tumor necrosis factor-alpha (TNF-M-NM-1), and bone morphogenetic proteins (BMP) and analyzed gene expression. cultured human periodontal ligament cells (control) VS cryopreserved and cultured periodontal ligament cells(cryopreserved group): 3 control replicates, 3 cryopreserved replicates
2013-01-19 | E-GEOD-43611 | biostudies-arrayexpress
Project description:Having the correct number of chromosomes is vital for normal development and health. Sex chromosome trisomy affects 0.1% of the human population and is associated with infertility. We show that during reprogramming to induced pluripotent stem cells (iPSCs), fibroblasts from sterile trisomic XXY and XYY mice lose the extra sex chromosome through a phenomenon we term trisomy-biased chromosome loss (TCL). Resulting euploid XY iPSCs can be differentiated into the male germ cell lineage and functional sperm that can be used in intracytoplasmic sperm injection to produce chromosomally normal, fertile offspring. Sex chromosome loss is comparatively infrequent during mouse XX and XY iPSC generation. TCL also applies to other chromosomes, generating euploid iPSCs from cells of a Down syndrome mouse model. It can also create euploid iPSCs from human trisomic patient fibroblasts. The findings have relevance to overcoming infertility and other trisomic phenotypes.
| S-EPMC5581950 | biostudies-literature
Project description:Cryoinjury and protein changes are a consequence of cryopreservation and may have a negative impact on sperm quality regarding motility, viability and fertilizing ability. However, potential proteomic changes in rabbit semen throughout the cryopreservation process have never been investigated previously. The aim of the present study was to compare the whole proteome of fresh and cryopreserved rabbit extracellular fluid of semen. Comparative analysis and identification of proteins were performed using 2-dimensional difference in-gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption/ionization mass (MALDI TOF/TOF) spectrometry. We found that 28 proteins differed in abundance between fresh and frozen extracellular fluid.
2022-10-13 | PXD034853 | Pride
Project description:BackgroundMany research laboratories have long-term repositories of cryopreserved peripheral blood mononuclear cells (PBMC), which are costly to maintain but are of uncertain utility for immunological studies after decades in storage. This study investigated preservation of cell surface phenotypes and in-vitro functional capacity of PBMC from viraemic HIV+ patients and healthy seronegative control subjects, after more than 20 years of cryopreservation.MethodsPBMC were assessed by 18-colour flow cytometry for major lymphocyte subsets within T, B, NK, and dendritic cells and monocytes. Markers of T-cell differentiation and activation were compared with original immunophenotyping performed in 1995/1996 on fresh blood at the time of collection. Functionality of PBMC was assessed by culture with influenza antigen or polyclonal T-cell activation, to measure upregulation of activation-induced CD25 and CD134 (OX40) on CD4 T cells and cytokine production at day 2, and proliferative CD25+ CD4 blasts at day 7. RNA was extracted from cultures containing proliferating CD4+ blast cells, and intracellular HIV RNA was measured using short amplicons for both the Double R and pol region pi code assays, whereas long 4-kbp amplicons were sequenced.ResultsAll major lymphocyte and T-cell subpopulations were conserved after long-term cryostorage, except for decreased proportions of activated CD38+HLA-DR+ CD4 and CD8 T cells in PBMC from HIV+ patients. Otherwise, differences in T-cell subpopulations between recent and long-term cryopreserved PBMC primarily reflected donor age-associated or HIV infection-associated effects on phenotypes. Proportions of naïve, memory, and effector subsets of T cells from thawed PBMC correlated with results from the original flow cytometric analysis of respective fresh blood samples. Antigen-specific and polyclonal T-cell responses were readily detected in cryopreserved PBMC from HIV+ patients and healthy control donors. Intracellular HIV RNA quantitation by pi code assay correlated with original plasma viral RNA load results. Full-length intracellular and supernatant-derived amplicons were generated from 5/12 donors, and sequences were ≥80% wild-type, consistent with replication competence.ConclusionsThis unique study provides strong rationale and validity for using well-maintained biorepositories to support immunovirological research even decades after collection.
| S-EPMC11361978 | biostudies-literature
Project description:Cryoinjury and protein changes are a consequence of cryopreservation and may have a negative impact on sperm quality regarding motility, viability and fertilizing ability. However, potential proteomic changes in rabbit semen throughout the cryopreservation process have never been investigated previously. The aim of the present study was to compare the whole proteome of fresh and cryopreserved rabbit spermatozoa. Comparative analysis and identification of proteins were performed using 2-dimensional difference in-gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption/ionization mass (MALDI TOF/TOF) spectrometry. We found that 108 proteins differed in abundance in spermatozoa between fresh and frozen semen. Most of these proteins was involved in pathways related to energy metabolism and protein quality control under stress conditions, reproductive processes and mechanisms of cell death/survival regulation, resulting in a significant decrease of motility and viability of post-thawing rabbit sperm and its potential fertilizing ability.
2022-10-14 | PXD034832 | Pride
Project description:The data in this study were obtained from proteomics and phosphoproteomics analyses of bull sperm during the cryopreservation process, with or without the addition of melatonin. The experiment employed TMT-10 plex labeling technology to analyze the proteome and phosphoproteome of three groups: before sperm cryopreservation, after cryopreservation without melatonin, and after cryopreservation with melatonin.
2024-10-23 | PXD057125 |