Ontology highlight
ABSTRACT:
PROVIDER: PRJNA753988 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR15430800.fastq.gz | Fastqsanger.gz | |||
| SRR15430801.fastq.gz | Fastqsanger.gz | |||
| SRR15430802.fastq.gz | Fastqsanger.gz | |||
| SRR15430803.fastq.gz | Fastqsanger.gz | |||
| SRR15430804.fastq.gz | Fastqsanger.gz |
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Project description:Cryopreservation of gametes offers significant potential for genetic diversity conservation and selective breeding in aquaculture. The blue mussel, Mytilus galloprovincialis, is valuable in the aquaculture industry, but declining seed availability and the need for efficient hatchery production, especially outside its natural reproductive season, make gamete cryopreservation essential. However, cryopreserving gametes, particularly oocytes from marine invertebrates, is challenging due to the balance required between cryoprotectant cytotoxicity and cell protection during freezing-thawing. This study investigates the effects of two common cryoprotectants, dimethyl sulfoxide (DMSO) and ethylene glycol (EG), on M. galloprovincialis oocytes using a monitored slow freezing (MSF) protocol. Our integrative analytical approach combined proteomics, functional, and ultrastructural analyses. Findings highlight that cryoprotectants induce significant proteomic alterations, more pronounced with DMSO than EG. These alterations are linked to oxidative stress, ineffective antioxidant response, and disruptions in meiosis restart mechanisms, leading to delayed larvae development. The MSF protocol resulted in plasma membrane rupture and lack of fertilisation success post-thawing. Our results provide the first report on the molecular basis underlying the limited success in cryopreserving mussel oocytes, showing EG as less harmful compared to DMSO, and supports supplementing cryoprotectants with external antioxidants and membrane stabilisers to improve cryopreservation of marine invertebrate oocytes.
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Project description:This study investigates the impact of cryopreservation and freezing media on RNA quality and gene expression profiles of peripheral blood mononuclear cells (PBMC). No significant difference in cell viability or RNA quality was observed between freshly isolated and cryopreserved cells, even after two freeze-thaw cycles. Transcriptome analysis revealed no significant differences in alignment rates or read counts across various conditions. Differential gene expression analysis identified changes between fresh and thawed samples, with a consistent downward trend of mRNA abundance in the frozen samples, but minimal changes were observed between the first and second freeze-thaw cycles. Gene set enrichment analysis showed only minor significant differences, and stress-related gene sets were not upregulated. Freezing medium appears to be the safer option compared to freezing in trizol. These results confirm that cryopreservation and thawing processes do not compromise RNA integrity or sequencing reliability in PBMC, supporting the use of cryopreserved samples for gene expression studies.
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Project description:After thawing, the mechanism of apoptosis advancing in human ES cells was invesitigated mainly by Gene conpath analysis. As time passed after thawing, nuclear fragmented cells increased more and more in cR-/mR- and cR+/mR-, but in cR+/mR+ and cR-/mR+, thawed cells formed cluster markedly, which was seen among three cell lines. Flow cytometry using TUNEL assay supported final colony formation ratio, which defined that such cell death was due to apoptosis. Gene map analysis 12 hours later after thawing of cR-/mR- showed activation of IL-1? & its receptor IL-1R and TGF-? & its receptor ACVR1C required for activation of caspase-8, initiation of caspase-8 and 10 and marked activation of ARHGDIB promoting actin reorganization comparing to cR+/mR+. Recovered cell lines in any condition did not lose proliferation and pluripotency regardless of ROCK inhibitor treatment.?These results showed apoptotic events after thawing were more marked in cR- and advanced through autoenhanced cascaded mechanism followed by initiation of self-cytokain activity, and were suppressed by ROCK inhibitor Y-27632. To investigate the protective mechanism caused by a novel cryo-technique of human embryonic stem (hES) cells using ROCK inhibitor Y-27632 against the cell death due to cryopreservation and thawing with Gene map analysis in addition to estimations of survival rate, apoptosis, undifferentiated states and pluripotency, the dissociated hES cells were cryopreserved in a freezing container in the following four conditions: addition of 10µM of Y-27632 to 1: cryoprotection solution and post-thawing medium (cR+/mR+), 2: only addition to cryoprotection (cR+/mR-), 3: only addition to post-thawing medium (cR-/mR+), 4: no addition to any medium (cR-/mR-).
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Project description:Some biosamples obtained from biobanks may go through thawing before processing. We aim to evaluate the effects of thawing at room temperature for different time periods on gene expression analysis.
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