NBDC Research ID: hum0490.v1
SUMMARY
Aims: In recent years, the Progestin-Primed Ovarian Stimulation (PPOS) method, utilizing progestin as ovulation suppressants, has gained traction as a controlled ovarian stimulation strategy in assisted reproductive technology. However, the impact of progestin agents on follicular fluid, mural granulosa cell (mGC), and cumulus cells at the levels of gene and protein expression has not been fully assessed. This study aims to evaluate the effects of progestin agents used in the PPOS method, on follicular development, and oocyte maturation.
Methods: Collected and washed mGCs were cryopreserved in Bambanker medium at −80 °C. After thawing, cell counts were standardised across patients and normalised to those of the patient with the minimum cell count. Cells were mixed within each group to form the PPOS and GnRH-ant groups. Dead cells were removed through magnetic cell sorting (Dead Cell Removal Kit, Miltenyi Biotec, Bergisch Gladbach, Germany) with LS columns and pre-separation filters from the MidiMACS™ Starting Kit (Miltenyi Biotec), following DNase treatment. After library preparation using Chromium Next GEM Single Cell V(D)J Reagent Kits, single cell RNA-seq was performed using a DNBSEQ-G400 sequencer.
Participants/Materials: mGCs from 16 infertility patients with normal ovarian function. Eight patients were treated with conventional ovulation induction (GnRH-ant) and eight patients were treated with PPOS.
| Dataset ID | Type of Data | Criteria | Release Date |
|---|---|---|---|
| JGAS000770 | NGS (scRNA-seq) | Controlled-access (Type I) | 2024/12/18 |
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MOLECULAR DATA
| Participants/Materials |
infertility patients with normal ovarian function (ICD10: N97): 16 cases - treated with GnRH-ant: 8 cases - treated with PPOS: 8 cases |
| Targets | scRNA-seq |
| Target Loci for Capture Methods | - |
| Platform | MGI [DNBSEQ-G400] |
| Library Source | RNAs extracted from granulosa cells which mixed within each group to form the GnRH-ant and PPOS |
| Cell Lines | - |
| Library Construction (kit name) | Chromium Next GEM Single Cell V(D)J Reagent Kits |
| Fragmentation Methods | Enzymatic fragmentation (10x Genomics Kit) |
| Spot Type | Paired-end |
| Read Length (without Barcodes, Adaptors, Primers, and Linkers) | 72 bp (R1), 91 bp (R2) |
| Mapping Methods | 10x Genomics Cell Ranger Count |
| Mapping Quality | Reads Mapped to Genome 98.9% (TFHS000278), 98.6% (TFHS000279) |
| Reference Genome Sequence | hg38 |
| Japanese Genotype-phenotype Archive Dataset ID | JGAD000911 |
| Total Data Volume | 126.3 GB (bam) |
| Comments (Policies) | NBDC policy |
DATA PROVIDER
Principal Investigator: Mika Handa
Affiliation: Obstetrics and Gynecology, Osaka University
Project / Group Name: -
Funds / Grants (Research Project Number):
| Name | Title | Project Number |
|---|---|---|
| KAKENHI Grant-in-Aid for Scientific Research (C) | Elucidating the Mechanism of Progestin Effects on Oocytes and Establishing an Optimal Treatment Protocol for the PPOS Method | 23K08867 |
PUBLICATIONS
| Title | DOI | Dataset ID | |
|---|---|---|---|
| 1 | Adverse effects of progestin-primed ovarian stimulation: Combination of clinical study and single cell analysis (under review) | JGAD000911 | |
| 2 |
USRES (Controlled-access Data)
| Principal Investigator | Affiliation | Country/Region | Research Title | Data in Use (Dataset ID) | Period of Data Use |
|---|---|---|---|---|---|
