1. Ezzatollah Fathi1, Raheleh Farahzadi 2, *, Najmeh Sheikhzadeh3
COMPARISION OF MULTIPOTENCY AND PLURIPOTENCY POTENTIAL IN ZEBRAFISH MESENCHYMAL STEM CELLS FROM HEART AND LIVER TISSUE
Ezzatollah Fathi1, Raheleh Farahzadi 2, *, Najmeh Sheikhzadeh3
1 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.
2 Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
3 Department of Food Hygiene and Aquatic Animals, Faculty of Veterinary Medicine, University of
Tabriz, Tabriz, Iran.
*Corresponding Author: Raheleh Farahzadi, PhD ( postal code: ; Tel: ; Fax: ; Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran).
Objectives
Methods
Results
There has been an explosion of research, publications and conference in the field of stem cells in the last decade. Mesenchymal stem cells (MSCs) are undifferentiated cells found in many adult tissues, including adipose, bone marrow, brain, muscle, ocular, heart, liver etc. In addition, MSCs have been considered as an appropriate source for gene and cell therapy tools for tissue engineering and regenerative medicine. To clarify different aspects of MSCs biology and develop MSCs applications in human diseases treatment, other studies on animal models are required. However, the similarity between zebrafish and human genome has been further examined; among other animal models that have been used, zebrafish is considered to be the best for practical purposes in regenerative medicine. In this report, multi-potency and pluri-potency potential of heart and liver derived-MSCs was presented for the first time. The aim of this study was to investigate the multipotency and pluripotency of zebrafish heart and liver derived-MSCs for using zebrafish as suitable animal model for regenerative medicine in cell therapy.
In the present study, MSCs were isolated from heart and liver tissue of Zebrafish (Danio rerio) as previously reported in another papers by Fathi et al. (2017). The flow-cytometry as well as RT-PCR were used to analyze the expression of a panel of cell surface markers CD44, CD90, CD31 and CD34. In the following, alizarin red, oil Red-O and toluidine blue staining were carried out to evaluate the multipotency of zebrafish heart and liver tissue-derived MSCs. Subsequently, the gene expression of Oct4, Sox2 and Nanog as pluri-potent markers were analyzed by RT-PCR.
The results showed that, like other MSCs, zebrafish heart and liver tissue-derived MSCs were expressed pluripotent markers Oct4, Sox2 and Nanog. Moreover, these cells were differentiated to osteocyte, adipocyte and chondrocyte lineages following directed differentiation. However, the findings of this study are consistent with the hypothesis that MSCs have similar features with other MSCs in different tissue.
Figure caption. The gene expression of cell surface markers for the heart (2C) and liver tissues (2D). 2E and F; gene expression profile of pluripotent markers from zebrafish heart and liver tissue-derived MSCs, respectively.
Conclusions
Our results show that both heart and liver tissue have similar characteristics including multi-lineage differentiation capacity and pluripotency potential.
Figure caption. In vitro multi-lineage differentiation of zebrafish heart and liver tissue-derived MSCs. (A) Osteogenic differentiation and alizarin red staining of mineralized cell aggregates; detection of the two bone specific genes (ALP and OCN) by RT-PCR method; (B) Generation of lipid vacuoles after adipogenesis; expression of PPAR-α and PPAR-γ as fat-specific genes; (C) After chondrogenesis of hADSCs, proteoglycan aggregates stain with toluidine blue; RT-PCR analysis of aggrecan and collagen II as chondrocyte-specific genes (bar = 40X).
References