PMID- 35055072
OWN - NLM
STAT- MEDLINE
DCOM- 20220207
LR  - 20220207
IS  - 1422-0067 (Electronic)
IS  - 1422-0067 (Linking)
VI  - 23
IP  - 2
DP  - 2022 Jan 14
TI  - Validating Reference Gene Expression Stability in Human Ovarian Follicles, 
      Oocytes, Cumulus Cells, Ovarian Medulla, and Ovarian Cortex Tissue.
LID - 10.3390/ijms23020886 [doi]
LID - 886
AB  - Human ovarian cells are phenotypically very different and are often only 
      available in limited amounts. Despite the fact that reference gene (RG) 
      expression stability has been validated in oocytes and other ovarian cells from 
      several animal species, the suitability of a single universal RG in the different 
      human ovarian cells and tissues has not been determined. The present study aimed 
      to validate the expression stability of five of the most used RGs in human 
      oocytes, cumulus cells, preantral follicles, ovarian medulla, and ovarian cortex 
      tissue. The selected genes were glyceraldehyde 3-phosphate dehydrogenase (GAPDH), 
      beta-2-microglobulin (B2M), large ribosomal protein P0 (RPLP0), beta-actin 
      (ACTB), and peptidylprolyl isomerase A (PPIA). Overall, the stability of all RGs 
      differed among ovarian cell types and tissues. NormFinder identified ACTB as the 
      best RG for oocytes and cumulus cells, and B2M for medulla tissue and isolated 
      follicles. The combination of two RGs only marginally increased the stability, 
      indicating that using a single validated RG would be sufficient when the 
      available testing material is limited. For the ovarian cortex, depending on 
      culture conditions, GAPDH or ACTB were found to be the most stable genes. Our 
      results highlight the importance of assessing RGs for each cell type or tissue 
      when performing RT-qPCR analysis.
FAU - Cadenas, Jesus
AU  - Cadenas J
AUID- ORCID: 0000-0001-8164-3697
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
FAU - Pors, Susanne Elisabeth
AU  - Pors SE
AUID- ORCID: 0000-0003-1546-1945
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
FAU - Nikiforov, Dmitry
AU  - Nikiforov D
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
FAU - Zheng, Mengxue
AU  - Zheng M
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
FAU - Subiran, Cristina
AU  - Subiran C
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
FAU - Botkjaer, Jane Alro
AU  - Botkjaer JA
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
FAU - Mamsen, Linn Salto
AU  - Mamsen LS
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
FAU - Kristensen, Stine Gry
AU  - Kristensen SG
AUID- ORCID: 0000-0003-1194-8393
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
FAU - Andersen, Claus Yding
AU  - Andersen CY
AD  - Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children 
      and Reproduction, University Hospital of Copenhagen, Rigshospitalet, 2100 
      Copenhagen, Denmark.
LA  - eng
GR  - 0134-00448/Independent Research Fund Denmark/
PT  - Journal Article
DEP - 20220114
PL  - Switzerland
TA  - Int J Mol Sci
JT  - International journal of molecular sciences
JID - 101092791
RN  - 0 (Biomarkers)
SB  - IM
MH  - *Biomarkers
MH  - Cumulus Cells/*metabolism
MH  - Female
MH  - Gene Expression Profiling
MH  - *Gene Expression Regulation
MH  - Humans
MH  - Oocytes/*metabolism
MH  - Ovarian Follicle/*metabolism
MH  - Ovary/*metabolism
MH  - RNA Stability
MH  - Transcriptome
PMC - PMC8778884
OTO - NOTNLM
OT  - NormFinder
OT  - housekeeping genes
OT  - human oocytes
OT  - ovarian tissue
OT  - preantral follicles
OT  - quantitative real-time PCR
COIS- The authors declare no conflict of interest.
EDAT- 2022/01/22 06:00
MHDA- 2022/02/08 06:00
PMCR- 2022/01/14
CRDT- 2022/01/21 01:10
PHST- 2021/12/17 00:00 [received]
PHST- 2022/01/08 00:00 [revised]
PHST- 2022/01/11 00:00 [accepted]
PHST- 2022/01/21 01:10 [entrez]
PHST- 2022/01/22 06:00 [pubmed]
PHST- 2022/02/08 06:00 [medline]
PHST- 2022/01/14 00:00 [pmc-release]
AID - ijms23020886 [pii]
AID - ijms-23-00886 [pii]
AID - 10.3390/ijms23020886 [doi]
PST - epublish
SO  - Int J Mol Sci. 2022 Jan 14;23(2):886. doi: 10.3390/ijms23020886.