Evaluation of β-actin as a Reference Gene for Comparative Expression Analysis of Equine Adipose- and Bone Marrow-Derived Mesenchymal Stem Cells by qRT-PCR

Authors

  • Abbas Parham Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine & Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
  • Adham Fani Maleki Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine & Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
  • Fatemeh Nazari Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine & Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Abstract:

Background Bone marrow and adipose tissue are two main sources of mesenchymal stem cells (MSCs). Some of studies suggest that there are some differences in gene expression profile of MSCs-derived from various tissues. To investigate gene expression profile by qRT-PCR, an appropriate reference gene with stable expression level should be chosen for normalizing data.  This study was designed to evaluate the stability of β-actin expression as a reference gene for studying comparative gene expression analysis of equine adipose- and marrow-derived MSCs. Materials and Method: MSCs were isolated from adipose tissue and bone marrow of two mares and cultured until passage 3 (P3). Total RNA of P3 cells was extracted and purity and quantity of RNA was assessed. cDNA was synthesized and qRT-PCR was performed in triplicate with β-actin primers. Results: Our analyses indicated that expression level of β-actin gene is different between adipose- and marrow-derived MSCs significantly. Mean ± SD of Ct was 21.13 ± 0.96 and 16.02 ± 0.88   for bone marrow- and adipose derived MSCs, respectively. Conclusion: Based on the results, it is suggested that β-actin is not a suitable gene for comparative gene expression analyses of equine adipose- and marrow- derived MSCs. Keywords: Adipose, Bone marrow, Equine, β-actin, Mesenchymal stem cells.  

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Journal title

volume 2  issue 2.3

pages  91- 91

publication date 2014-05-01

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