Comparison of the liver function and hepatic specific genes expression in cultured mesenchymal stem cells and hepatocytes

Document Type : Original Article

Authors

1 School of Biology, Damghan University, Damghan, Iran

2 School of Biology, Damghan University, Damghan, Iran 2 Institute of Biological Sciences, Damghan University, Damghan, Iran

Abstract

Objective(s):
Stem cell therapy is believed to be as a promising treatment strategy for tissue repair and regeneration. The plasticity specification of the adult stem cells, such as MSCs, has enabled that these cells to be used in the treatment of a broad spectrum of diseases like liver disorders. In this study, the production of urea and Albumin (Alb), glycogen storage, and expression of some liver genes including α
-fetoprotein (AFP), Alb, cytokeratin18 (CK18) and cytokeratin19 (CK19) was compared between mesenchymal stem cells (MSCs) and isolated rat hepatocytes.
 Materials and Methods:
The MSCs were isolated from rat femurs and tibias and cultured in α
-MEM, DMEM and RPMI mediums supplemented with serum. Hepatocytes were isolated from Rat livers and cultured in DMEM with serum. The expression of AFP, Alb, CK18, and CK19 genes was evaluated using the reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, the synthesis of albumin and urea of the cells was measured
 Results:
 In vitro
conditions, MSCs and hepatocytes exhibited the characteristic functions of the liver such as capacity to synthesize Alb, urea, the storage of glycogen. In this study, the expression of some liver genes such as AFP, Alb, CK18 and CK19 at mRNA levels was also shown.
 
 

Keywords

References

1. Kallis YN, Alison MR, Forbes SJ. Bone marrow stem cells and liver disease. Gut. 2007; 56:716-724.
2. Oertel M, Shafritz DA. Stem cells, cell transplantation and liver repopulation. Biochim Biophys Acta. 2008; 1782:61-74.
3. Cantz T, Manns MP, Ott M. Stem cells in liver regeneration and therapy. Cell Tissue Res. 2008; 331:271-282.
4. Seki T, Yokoyama Y, Nagasaki H, Kokuryo T, Nagino M. Adipose tissue-derived mesenchymal stem cell transplantation promotes hepatic regeneration after hepaticischemia-reperfusion and subsequent hepatectomy in rats. J Surg Res. 2012; 178:63-70.
5. Almeida-Porada G, Zanjani ED, Porada CD. Bone marrow stem cells and liver regeneration. Exp Hematol. 2010; 38:574-580.
6. Fürst G, Schulte am Esch J, Poll LW, Hosch SB, Fritz LB, Klein M,
et al
. Portal vein embolization and autologous CD133+ bone marrow stem cells for liver regeneration: initial experience. Radiology. 2007; 243:171-179.
7. Kanazawa H, Fujimoto Y, Teratani T, Iwasaki J, Kasahara N, Negishi K,
 
 
et al
. Bone marrow-derived
 
Nikoozad et al Liver function and hepatic gene expression
Iran J Basic Med Sci, Vol. 17, No. 1, Jan 2014
11
mesenchymal stem cells ameliorate hepatic ischemia reperfusion injury in a rat model. PLoS One. 2011; 6:e19195.
8. Dai LJ, Li HY, Guan LX, Ritchie G, Zhou JX. The therapeutic potential of bone marrow-derived mesenchymal stem cells on hepatic cirrhosis. Stem Cell Res. 2009; 2:16-25.
9. Cho KA, Ju SY, Cho SJ, Jung YJ, Woo SY, Seoh JY,
et al
. Mesenchymal stem cells showed the highest potential for the regeneration of injured liver tissue compared with other subpopulations of the bone marrow. Cell Biol Int. 2009; 33:772-777.
10. Qihao Z, Xigu C, Guanghui C, Weiwei Z. Spheroid formation and differentiation into hepatocyte-like cells of rat mesenchymal stem cell induced by co-culture with liver cells. DNA Cell Biol. 2007; 26:497-503.
 
11. Mohsin S, Shams S, Ali Nasir G, Khan M, Javaid Awan S, Khan SN,
 
 
et al
. Enhanced hepatic differentiation of mesenchymal stem cells after pretreatment with injured liver tissue. Differentiation. 2011; 81:42-48.
12. Azizi SA, Stokes D, Augelli BJ, DiGirolamo C, Prockop DJ. Engraftment and migration of human bone marrow stromal cells implanted in the brains of albino rats--similarities to astrocyte grafts. Proc Natl Acad Sci U S A. 1998; 95:3908-3913.
 
13. Davis JM. Basic cell culture. New York, NY: Oxford University Press; 2002.
 
14. Dutta A, Bandyopadhyay S, Mandal C, Chatterjee M. Development of a modified MTT assay for screening antimonial resistantfield isolates of Indian visceral leishmaniasis. Parasitol Int. 2005; 54:119-122.
 
15. Xie C, Zheng YB, Zhu HP, Peng L, Gao ZL. Human bone marrow mesenchymal stem cells are resistant to HBV infection during differentiation into hepatocytes
 
 
in vivo and in vitro
. Cell Biol Int. 2009; 33:493-500.
16. Taghi GM, Ghasem Kashani Maryam H, Taghi L, Leili H, Leyla M. Characterization of
 
 
in vitro
cultured bone marrow and adipose tissue-derived mesenchymal stem cells and their ability to express neurotrophic factors. Cell Biol Int. 2012; 36:1239-1249.
17. Stöcker E, Wullstein HK, Bräu G. [Capacity of regeneration in liver epithelia of juvenile, repeated partially hepatectomized rats. Autoradiographic studies after continous infusion of 3H-thymidine (author's transl)]. Virchows Arch B Cell Pathol. 1973; 14:93-103.
 
18. Dalakas E, Newsome PN, Harrison DJ, Plevris JN. Hematopoietic stem cell trafficking in liver injury. FASEB J. 2005; 19:1225-1231.
 
19. Miyazaki M, Akiyama I, Sakaguchi M, Nakashima E, Okada M, Kataoka K,
 
 
et al
. Improved conditions to induce hepatocytes from rat bone marrow cells in culture. Biochem Biophys Res Commun. 2002; 298:24-30.
20. Nakamura T, Nishizawa T, Hagiya M, Seki T, Shimonishi M, Sugimura A,
 
 
et al
. Molecular cloning and expression of human hepatocyte growth factor. Nature. 1989; 342:440-443.
21. Nakamura T, Teramoto H, Ichihara A. Purification and characterization of a growth factor from rat platelets for mature parenchymal hepatocytes in primary cultures. Proc Natl Acad Sci USA. 1986; 83: 6489-6493.
 
22. Gohda E, Tsubouchi H, Nakayama H, Hirono S, Sakiyama O, Takahashi K,
 
 
et al
. Purification and partial characterization of hepatocyte growth factor from plasma of a patient with fulminant hepatic failure. J Clin Invest. 1988; 81:414-419.
23. Lee KD, Kuo TK, Whang-Peng J, Chung YF, Lin CT, Chou SH,
 
 
et al. In vitro
hepatic differentiation of human mesenchymal stem cells. Hepatology. 2004; 40:1275-1284.
24. Baksh D, Song L, Tuan RS. Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med. 2004; 8:301-316.
 
25. Sethe S, Scutt A, Stolzing A. Aging of mesenchymal stem cells. Ageing Res Rev. 2006; 5:91-116.
 
26. Pountos I, Corscadden D, Emery P, Giannoudis PV. Mesenchymal stem cell tissue engineering: Techniques for isolation, expansion and application. Injury. 2007; 38:S23-33.
 
27. Meirelles Lda S, Fontes AM, Covas DT, Caplan AI. Mechanisms involved in the therapeutic properties of mesenchymal stem cells. Cytokine Growth Factor Rev. 2009; 20:419-427.
 
28. Baddoo M, Hill K, Wilkinson R, Gaupp D, Hughes C, Kopen GC,
 
 
et al
. Characterization of mesenchymal stem cells isolated from murine bone marrow by negative selection. J Cell Biochem. 2003; 89:1235-1249.
29. Chen Y, Shao JZ, Xiang LX, Dong XJ, Zhang GR. Mesenchymal stem cells: A promising candidate in regenerative medicine. Int J Biochem Cell Biol. 2008; 40:815-820.
 
30. Dai LJ, Li HY, Guan LX, Ritchie G, Zhou JX. The therapeutic potential of bone marrow-derived mesenchymal stem cells on hepatic cirrhosis. Stem Cell Res. 2009; 2:16-25.
 
31. Luk JM, Wang PP, Lee CK, Wang JH, Fan ST. Hepatic potential of bone marrow stromal cells: Development of
 
 
in vitro
co-culture and intra-portal transplantation models. J Immunol Methods. 2005; 305:39-47.
32. Wang YJ, Liu HL, Guo HT, Wen HW, Liu J. Primary hepatocyte culture in collagen gel mixture and collagen sandwich. World J Gastroenterol. 2004; 10:699-702.
 
33. Zhao R, Duncan SA. Embryonic development of the liver. Hepatology. 2005; 41: 956-967.
 
34. Shiojiri N, Lemire JM, Fausto N. Cell lineages and oval cell progenitors in rat liver development Cancer Res. 1991; 51:2611-2620.
 
35. Jakubowski A, Ambrose C, Parr M, Lincecum JM, Wang MZ, Zheng TS,
 
 
et al
. TWEAK induces liver progenitor cell proliferation. J Clin Invest. 2005; 115:2330-2340.
36. Gordon GJ, Coleman WB, Hixson DC, Grisham JW. Liver regeneration in rats with retrosin-induced hepatocellular injury proceeds through a novel cellular response. Am J Pathol. 2000; 156:607-619.
 
37. Mall FP. A study of the structural unit of the liver. Am J Anat. 1906; 5:227-308.
 
38. Rothenberg ME, Cali JJ, Sobol M, Briggs MW, Upton T. Rat Hepatocyte Culture Physiology Shows Enhanced Cytochrome P450 Activity on a Synthetic Extracellular Matrix.
 
 
In vitro
toxicology. 2008; 20:18-20.
39. Lysy PA, Campard D, Smets F, Najimi M, Sokal EM. Stem cells for liver tissue repair: current knowledge and perspectives. World J Gastroenterol. 2008; 14: 864-875.
 
40. Fausto N. Liver regeneration and repair: hepatocytes, progenitor cells, and stem cells. Hepatology. 2004; 39: 1477-1487.
 
41. Serralta A, Donato MT, Martinez A, Pareja E, Orbis F, Castell JV,
et al
 

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