Level D· Scientific groundwork from lab and animal studiesAnimal StudyPubMedOpen access

Selection and Validation of Reference Genes for Gene Expression Studies in an Equine Adipose-Derived Mesenchymal Stem Cell Differentiation Model by Proteome Analysis and Reverse-Transcriptase Quantitative Real-Time PCR.

Riveroll AL., Skyba-Lewin S., Lynn KD., Mubyeyi G., Abd-El-Aziz A., Kibenge FST.

Animal Study, published in Genes (Basel) (2023) — summary generated from the PubMed abstract.

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Level D· Scientific groundwork from lab and animal studiesEvidence level of this study

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

  • Level A · Stronger Clinical Evidence
  • Level B · Emerging clinical evidence with positive signals
  • Level C · Early human research exploring benefits
  • Level D · Scientific groundwork from lab and animal studies
  • Emerging · Emerging topic under active research
Read the A–D evidence level guide

This page is generated from the PubMed record. The Thai description is an automated summary of bibliographic fields and the abstract, not a full translation, and is not medical advice.

Study type
Animal Study
Journal
Genes (Basel) (2023)
Country
Switzerland
Reported sample size
—
Source database
PubMed
PMID
36980946
PMCID
PMC10048155
DOI
10.3390/genes14030673
Citations
5

Abstract (original English)

Adipose-derived stem cells (ADSCs) are used in tissue regeneration therapies. The objective of this study is to identify stable reference genes (RGs) for use in gene expression studies in a characterized equine adipose-derived mesenchymal stem cell (EADMSC) differentiation model. ADSCs were differentiated into adipocytes (ADs) or osteoblasts (OBs), and the proteomes from these cells were analyzed by liquid chromatography tandem mass spectrometry. Proteins that were stably expressed in all three cells types were identified, and the mRNA expression stabilities for their corresponding genes were validated by RT-qPCR. PPP6R1 , CCDC97 , and then either ACTB or EPHA2 demonstrated the most stable mRNA levels. Normalizing target gene C q data with at least three of these RGs simultaneously, as per MIQE guidelines ( PPP6R1 and CCDC97 with either ACTB or EPHA2 ), resulted in congruent conclusions. FABP5 expression was increased in ADs (5.99 and 8.00 fold, p = 0.00002 and p = 0.0003) and in OBs (5.18 and 5.91 fold, p = 0.0011 and p = 0.0023) relative to ADSCs. RUNX2 expression was slightly higher in ADs relative to ADSCs (1.97 and 2.65 fold, p = 0.04 and p = 0.01), but not in OBs (0.9 and 1.03 fold, p = 0.58 and p = 0.91).

What this study does not prove

  • • This study does not prove SVF is an approved treatment or a replacement for standard care.
  • • This is preclinical work; animal or laboratory results cannot be applied to humans.

Evidence level

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

How we grade evidence
AnimalsHorsesProteomeRNA-Directed DNA PolymeraseReal-Time Polymerase Chain ReactionCell DifferentiationMesenchymal Stem CellsGene ExpressionRNA, MessengerDNA-Directed RNA Polymerases

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