ISOLATION AND CHARACTERIZATION OF FETAL AND NEONATAL LIVER-DERIVED MESENCHYMAL STEM CELLS FROM RABBITS

Authors

  • Esraa Elsheikh Department of Anatomy and Embryology, Faculty of Veterinary Medicine, Zagazig University. Alzeraa Street, Postal code 44511, Zagazig City, Al-Sharkia Province, Egypt
  • Enas EL-Hady 1Anatomy and Embryology Department, Faculty of Veterinary Medicine, Zagazig University
  • Somia Abdallah 2 Biochemistry and Molecular Biology Department, Faculty of Medicine, Zagazig University
  • Atef Selem 1Anatomy and Embryology Department, Faculty of Veterinary Medicine, Zagazig University
  • Mervat Konsowa 1Anatomy and Embryology Department, Faculty of Veterinary Medicine, Zagazig University

DOI:

https://doi.org/10.26873/SVR-1438-2021

Keywords:

liver, mesenchymal stem cells, immunohistochemistry, flow cytometry, rabbit

Abstract

Mesenchymal stem cells (MSCs) became in recent years a promising implement for cell therapies. Our study described the isolation and characterization of liver mesenchymal stem cells (LMSCs). We isolated LMSCs from the rabbit liver tissue from two groups: 20th day's old fetuses and 4 days-old kittens. LMSCs were characterized by flow cytometry, immunohistochemistry (IHC), and quantitative real-time-polymerase chain reaction (qRT-PCR). Flow cytometry of the isolated LMSCs showed variable expression of the cluster of differentiation markers (CD) including, CD105, CD106, CD11b, CD38, CD63, CD9, and CD14. Fetal liver isolated LMSCs showed expression of CD105, CD106, CD9, and CD63 but no expression of CD14, CD11b, and CD38 was observed. Absolute expression of LMSCs was performed by qRT-PCR analysis to detect cytoskeleton genes including, vimentin, nestin, and desmin in the experimental groups. In vivo localization of stem cell markers such as CD105 (Endoglin), vimentin, and desmin was also distinguished in fetal and neonatal liver tissues using IHC.

 

References

● 1. McGeady TA, Quinn PJ, FitzPatric ES, Ryan MT, Kilroy D, Lonergan P. Veteri-nary Embryology textbook. 2nd edition, Pondicherry, India, 2017; 59-66.

● 2. Saliem M, Ericzon B, Ellis E, Hovatta O, Götherström C. Isolation and Characteri-zation of Mesenchymal Stem Cells from Human Fetal Liver; Potential Candidates for Replacement Therapy in Liver Dis-ease. J Liver: Dis Transplant 2012; 1 (2): 1-9.

● 3. Moreno R, Gonzalez IM, Rosal M, Far-wati A, Gratacos E, Aran JM. Characteri-zation of Mesenchymal Stem Cells Iso-lated from the Rabbit Fetal Liver, STEM CELLS AND DEVELOPMENT 2010; 19 (10): 1579-1588.

● 4. Lotfy A, Abdelsamed M, Abdelrahman A, Nabil A, Abdel-Fatah A, Hozayen W, Shiha G. Markers for the Characterization of Liver Mesenchymal Stem Cell. Inter-national Journal of Cell Science & Mo-lecular Biology 2019; 6 (1): 116-121.

● 5. Curtis LAM, Wieczorek A J, McGann LE, Elliott JAW. Mesenchymal stromal cells derived from various tissues: Bio-logical, clinical and cryopreservation as-pects. Cryobiology 2015; 71(2):181-197.

● 6. Lu T, Hu P, Su X, Li C, Ma Y, Guan W. Isolation and characterization of mesen-chymal stem cells derived from fetal bo-vine liver. Cell Tissue Bank 2014; 15: 439–450.

● 7. Kobolak J, Dinnyes A, Memic A, Khademhosseini A, Mobasheri A. Mes-enchymal stem cells: Identification, phe-notypic characterization, biological prop-erties and potential for regenerative med-icine through biomaterial micro-engineering of their niche. Methods 2016; 99: 62–68.

● 8. Ullah I, Subbarao RB, Rho GJ. Human mesenchymal stem cells - current trends and future prospective. Biosci Rep 2015; 35 (2): 1-18.

● 9. Dan YY, Riehle KJ, Lazaro C, Teoh N, Haque J, Campbell JS, Fausto N. Isola-tion of multipotent progenitor cells from human fetal liver capable of differentiat-ing into liver and mesenchymal lineages. PNAS 2006; 103 (26): 9912–9917.

● 10. Turner R, Lozoya O, Wang Y, Cardi-nale V, Gaudio E, Alpini G, Mendel G, Wauthier E, Barbier C, Alvaro D, Reid LM. Human Hepatic Stem Cell and Mat-urational Liver Lineage Biology. Hepatol-ogy 2011; 53 (3): 1035–1045.

● 11. El-Kehdy H, Pourcher G, Zhang W, Hamidouche Z, Mainot SG, Sokal E, Charbord P, Najimi M, Kupperschmitt AD. Hepatocytic Differentiation Poten-tial of Human Fetal Liver Mesenchymal Stem Cells: In Vitro and In Vivo Evalua-tion. Stem Cells International 2016; 1-12.

● 12. Moreno R, Gonzalez IM, Rosal M, Nadal M, Petriz J, Gratacos E, Aran JM. Fetal Liver-Derived Mesenchymal Stem Cell Engraftment After Allogeneic In Utero Transplantation into Rabbits. Stem cells and development 2012; 21(2), 284-295.

● 13. Nikoozad Z, Ghorbanian MT, Rezaei A. Comparison of the liver function and hepatic specific genes expression in cul-tured mesenchymal stem cells and hepatocytes. Iran J Basic Med Sci 2014; 17:27-33.

● 14. Lee TC, Lee TH, Huang YH, Chang NK, Lin YJ, Chien PWC, Yang WH, Lin MHC. Comparison of Surface Markers between Human and Rabbit Mesenchy-mal Stem Cells. PLOS ONE 2014; 9(11): 1-10.

● 15. Suzuki A, Zheng Y, Kaneko S, On-odera M, Fukao K, Nakauchi H,Taniguchi H. Clonal identification and characterization of self-renewing pluripo-tent stem cells in the developing liver. The Journal of Cell Biology 2002; 156 (1): 173–184.

● 16. Ghaneialvar H, Soltani L, Rahmani HR, Lotfi AS, Soleimani M. Characterization and Classification of Mesenchymal Stem Cells in Several Species Using Surface Markers for Cell Therapy Purposes, Ind J Clin Biochem 2018; 33(1): 46–52.

● 17. Theuerkauf I, Zhou H, Fischer HP. Immuno-

histochemical patterns of human liver si-nusoids under different conditions of pathologic perfusion. Virchows Arch 2001; 438:498–504.

● 18. Yu D, Zhuang L, Sun X, Chen J, Yao Y, Meng K, Ding Y. Particular distribu-tion and expression pattern of endoglin (CD105) in the liver of patients with hepatocellular carcinoma. BMC Cancer 2007; 7(122):1-10.

● 19. Gumerova AA, Titova MA, Kiassov AP. Expression of Different Liver Cell Markers during the Early Prenatal Hu-man Development. CELL AND Tissue biology 2007; 1(2): 143–150.

● 20. Kiassov AP, Eyken PV, Pelt JFV, Depla E,

Fevery J, Desmet VJ, Yap PSH. Desmin expressing non hematopoietic liver cells during rat liver development: An immu-no-histochemical and morphometric study. Differentiation 1995; 59:253-258.

● 21. Milijana K, Gledic D, Kukolj V, Knezevic DJ, Jovanovic M, Bozic T, San-ja AK. Expression of α-SMA, Desmin and Vimentin in Canine Liver with Fibro-sis. Acta Veterinaria (Beograd) 2009; 59 (4): 361-370.

● 22. Buyssens N, Kockx MM, Herman AG, Lazou JM, Berg KVD, Wisse E, Geerts A. Centrolobular Liver Fibrosis in the Hypercholesterolemic Rabbit. Hepatolo-gy 1996; 24(4): 939-946.

Downloads

Published

2021-12-17

How to Cite

Elsheikh, E., EL-Hady, E., Abdallah, S., Selem, A., & Konsowa, M. (2021). ISOLATION AND CHARACTERIZATION OF FETAL AND NEONATAL LIVER-DERIVED MESENCHYMAL STEM CELLS FROM RABBITS. Slovenian Veterinary Research, 58(24-Suppl), 177–86. https://doi.org/10.26873/SVR-1438-2021

Issue

Vol. 58 No. 24-Suppl (2021)

Section

Original Research Article

License

Copyright (c) 2021 Esraa Elsheikh, Enas EL-Hady, Somia Abdallah, Atef Selem, Mervat Konsowa

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Most read articles by the same author(s)