» Articles » PMID: 36009640

Comparison of Sources and Methods for the Isolation of Equine Adipose Tissue-Derived Stromal/Stem Cells and Preliminary Results on Their Reaction to Incubation with 5-Azacytidine

Overview
Journal Animals (Basel)
Date 2022 Aug 26
PMID 36009640
Authors
Affiliations
Soon will be listed here.
Abstract

Physiological particularities of the equine heart justify the development of an in vitro model suitable for investigations of the species-specific equine cardiac electrophysiology. Adipose tissue-derived stromal/stem cells (ASCs) could be a promising starting point from which to develop such a cardiomyocyte (CM)-like cell model. Therefore, we compared abdominal, retrobulbar, and subcutaneous adipose tissue as sources for the isolation of ASCs applying two isolation methods: the collagenase digestion and direct explant culture. Abdominal adipose tissue was most suitable for the isolation of ASCs and both isolation methods resulted in comparable yields of CD45-/CD34-negative cells expressing the mesenchymal stem cell markers CD29, CD44, and CD90, as well as pluripotency markers, as determined by flow cytometry and real-time quantitative PCR. However, exposure of equine ASCs to 5-azacytidine (5-AZA), reportedly inducing CM differentiation from rats, rabbits, and human ASCs, was not successful in our study. More precisely, neither the early differentiation markers and , nor the late CM differentiation markers , and were upregulated in equine ASCs exposed to 10 µM 5-AZA for 48 h. Hence, further work focusing on the optimal conditions for CM differentiation of equine stem cells derived from adipose tissue, as well as possibly from other origins, are needed.

Citing Articles

Influence of the Anatomical Site on Adipose Tissue-Derived Stromal Cells' Biological Profile and Osteogenic Potential in Companion Animals.

Ferreira-Baptista C, Ferreira R, Fernandes M, Gomes P, Colaco B Vet Sci. 2023; 10(12).

PMID: 38133224 PMC: 10747344. DOI: 10.3390/vetsci10120673.


Multilineage Differentiation Potential of Equine Adipose-Derived Stromal/Stem Cells from Different Sources.

Stage H, Trappe S, Sollig K, Trachsel D, Kirsch K, Zieger C Animals (Basel). 2023; 13(8).

PMID: 37106915 PMC: 10135324. DOI: 10.3390/ani13081352.

References
1.
Burk J, Ribitsch I, Gittel C, Juelke H, Kasper C, Staszyk C . Growth and differentiation characteristics of equine mesenchymal stromal cells derived from different sources. Vet J. 2012; 195(1):98-106. DOI: 10.1016/j.tvjl.2012.06.004. View

2.
Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D . Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006; 8(4):315-7. DOI: 10.1080/14653240600855905. View

3.
Yang J, Song T, Wu P, Chen Y, Fan X, Chen H . Differentiation potential of human mesenchymal stem cells derived from adipose tissue and bone marrow to sinus node-like cells. Mol Med Rep. 2011; 5(1):108-13. DOI: 10.3892/mmr.2011.611. View

4.
Lee D, Joo S, Han S, Im J, Lee S, Sonn C . Isolation and expansion of synovial CD34(-)CD44(+)CD90(+) mesenchymal stem cells: comparison of an enzymatic method and a direct explant technique. Connect Tissue Res. 2010; 52(3):226-34. DOI: 10.3109/03008207.2010.516850. View

5.
Kern S, Eichler H, Stoeve J, Kluter H, Bieback K . Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 2006; 24(5):1294-301. DOI: 10.1634/stemcells.2005-0342. View