Intrasplenic co-transplantation of fetal hepatic progenitor cells and transforming growth factor beta 1 induced hepatic stellate cells ameliorates acute liver injury

doi:10.3969/j.issn.2095-4344.0440

Abstract

Abstract:

BACKGROUND: Difficulty in long-time survival and continuous proliferation is the main problem for transplanted fetal hepatic progenitor cells and co-transplantation with transforming growth factor beta 1 (TGF-β1)-induced hepatic stellate cells may be a promising way to solve this scientific obstacle.
OBJECTIVE: To explore the therapeutic effects of co-transplantation of fetal hepatic progenitor cells and TGF-β1-induced hepatic stellate cells on acute liver injury in mice.
METHODS: Over-expression vector pHBLV-CMVIE-TGF-β1 was infected to mouse hepatic stellate cells and transfection efficiency was detected by immunocytochemistory, western blot and qRT-PCR. Hepatic progenitor cells, mHPCs-E_14.5, were cultured and identified by immunofluorescence in vitro. The mouse model of acute liver injury was established by intraperitoneal injection of CCl₄ in combination with 2/3 partial hepatectomy, followed by mHPCs-E_14.5 transplantation, co-transplantation of mHPCs-E_14.5 and mHSCs-pHBLV-CMVIE-TGF-β1 (experimental co-transplantation group) or co-transplantation of mHPCs-E_14.5 and mHSCs-pHBLV-CMVIE-GFP (control co-transplantation group) for cell transplantation assay. Confocal immunofluorescence staining against CK19, ALB, a-SMA was performed to analyze the engraftment and differentiation of transplanted cells in the splenic parenchyma 14 days post-transplantation; serum alanine transferase and aspartate transferase levels were monitored using an automatic biochemistry analyzer.
RESULTS AND CONCLUSION: (1) A hepatic stellate cell line that over-expressing TGF-β1 was successfully established and expression levels of TGF-β1 and α-smooth muscle actin were efficiently up-regulated in the over-expression group (P < 0.01). (2) mHPCs-E_14.5 expressed massive AFP and low levels of ALB and CK19, confirming that this cell line was in complete conformity with fetal hepatic progenitor cells in vitro. (3) CK19 and ALB positive cells existed in the splenic parenchyma in mHPCs-E_14.5 transplantation group. Highly expressed ALB but less expressed α-SMA and CK19 were observed in the control co-transplantation group, while massive CK19 and a-SMA positive cells as well as less level of ALB positive cells existed in the experimental co-transplantation group. Serum alanine transferase and aspartate transferase levels were decreased remarkably after cell transplantation, and moreover, the decrease was more obvious in the experimental co-transplantation group (P < 0.05). Overall, transplanted fetal hepatic progenitor cells engraft and differentiate into hepatocytes and cholangiocytes in the splenic parechyma successfully in vivo. Importantly, hepatic stellate cells induced by TGF-β1 promote the differentiation of fetal hepatic progenitor cells into cholangiocytes and accelerate recovery from CCl₄/partial hepatectomy induced acute liver injury.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Hepatic Stellate Cells, Stromal Cells, Liver Diseases, Cell Transplantation, Tissue Engineering

CLC Number:

R394.2

Aimaiti Yasen, Jin Xin, Chen Zi-xin, Li De-wei. Intrasplenic co-transplantation of fetal hepatic progenitor cells and transforming growth factor beta 1 induced hepatic stellate cells ameliorates acute liver injury[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(5): 710-716.

Figures/Tables 2

References

[1] Zeldenrust S, Gertz M, Uemichi T, et al. Orthotopic liver transplantation for hereditary fibrinogen amyloidosis. Transplantation. 2003;75(4):560-561.

[2] Rhim JA, Sandgren EP, Palmiter RD, et al. Complete reconstitution of mouse liver with xenogeneic hepatocytes. Proc Natl Acad Sci U S A. 1995;92(11):4942-4946.

[3] Pietrosi G, Vizzini GB, Gruttadauria S, et al. Clinical applications of hepatocyte transplantation. World J Gastroenterol. 2009;15(17):2074-2077.

[4] Ishii T, Yasuchika K, Machimoto T, et al. Transplantation of embryonic stem cell-derived endodermal cells into mice with induced lethal liver damage. Stem Cells. 2007;25(12): 3252-3260.

[5] Yap KK, Dingle AM, Palmer JA, et al. Enhanced liver progenitor cell survival and differentiation in vivo by spheroid implantation in a vascularized tissue engineering chamber. Biomaterials. 2013;34(16):3992-4001.

[6] Franco OE, Shaw AK, Strand DW, et al. Cancer associated fibroblasts in cancer pathogenesis. Semin Cell Dev Biol. 2010;21(1):33-39.

[7] Folkman J, Kalluri R. Cancer without disease.Nature. 2004; 427(6977):787.

[8] Gordillo M, Evans T, Gouon-Evans V. Orchestrating liver development. Development. 2015;142(12):2094-2108.

[9] Suzuki K, Tanaka M, Watanabe N, et al. p75 Neurotrophin receptor is a marker for precursors of stellate cells and portal fibroblasts in mouse fetal liver. Gastroenterology. 2008;135(1): 270-281.e3.

[10] Tanimizu N, Saito H, Mostov K, et al. Long-term culture of hepatic progenitors derived from mouse Dlk+ hepatoblasts. J Cell Sci. 2004;117(Pt 26):6425-6434.

[11] Cheng K, Benten D, Bhargava K, et al. Hepatic targeting and biodistribution of human fetal liver stem/progenitor cells and adult hepatocytes in mice. Hepatology. 2009;50(4):1194-1203.

[12] Chen L, Davis GJ, Crabb DW, et al. Intrasplenic transplantation of isolated periportal and perivenous hepatocytes as a long-term system for study of liver-specific gene expression. Hepatology. 1994;19(4):989-998.

[13] Martins PN, Theruvath TP, Neuhaus P. Rodent models of partial hepatectomies. Liver Int. 2008;28(1):3-11.

[14] Sancho-Bru P, Najimi M, Caruso M, et al. Stem and progenitor cells for liver repopulation: can we standardise the process from bench to bedside. Gut. 2009;58(4):594-603.

[15] Duncan AW, Dorrell C, Grompe M. Stem cells and liver regeneration. Gastroenterology. 2009;137(2):466-481.

[16] Zhang W, Tucker-Kellogg L, Narmada BC, et al. Cell-delivery therapeutics for liver regeneration. Adv Drug Deliv Rev. 2010; 62(7-8):814-826.

[17] Kalinichenko VV, Bhattacharyya D, Zhou Y, et al. Foxf1 +/- mice exhibit defective stellate cell activation and abnormal liver regeneration following CCl4 injury. Hepatology. 2003; 37(1): 107-117.

[18] Onitsuka I, Tanaka M, Miyajima A. Characterization and functional analyses of hepatic mesothelial cells in mouse liver development. Gastroenterology. 2010;138(4):1525-1535.

[19] Yasui O, Miura N, Terada K, et al. Isolation of oval cells from Long-Evans Cinnamon rats and their transformation into hepatocytes in vivo in the rat liver. Hepatology. 1997;25(2): 329-334.

[20] Weis SM, Cheresh DA. Tumor angiogenesis: molecular pathways and therapeutic targets. Nat Med. 2011;17(11): 1359-1370.

[21] Liu D, Xiao H, Du C, et al. The effect of fibroblast activation on vascularization in transplanted pancreatic islets. J Surg Res. 2013;183(1):450-456.

[22] Cai J, Ito M, Nagata H, et al. Treatment of liver failure in rats with end-stage cirrhosis by transplantation of immortalized hepatocytes. Hepatology. 2002;36(2):386-394.

[23] Gupta S, Vemuru RP, Lee CD, et al. Hepatocytes exhibit superior transgene expression after transplantation into liver and spleen compared with peritoneal cavity or dorsal fat pad: implications for hepatic gene therapy. Hum Gene Ther. 1994;5(8):959-967.

[24] Ahmad TA. Study of the survival of hepatocytes transplanted to the spleen in rats. Hepatogastroenterology. 2011;58 (107-108): 892-895.

[25] 万真,张晓刚,郑幸龙,等. 异种肝前体细胞移植治疗急性肝损伤大鼠[J].中国组织工程研究, 2013,17(53): 9132-9138.

[26] Pintilie DG, Shupe TD, Oh SH, et al. Hepatic stellate cells' involvement in progenitor-mediated liver regeneration. Lab Invest. 2010;90(8):1199-1208.

[27] Clotman F, Jacquemin P, Plumb-Rudewiez N, et al. Control of liver cell fate decision by a gradient of TGF beta signaling modulated by Onecut transcription factors. Genes Dev. 2005; 19(16):1849-1854.

[28] Zong Y, Stanger BZ. Molecular mechanisms of bile duct development. Int J Biochem Cell Biol. 2011;43(2):257-264.

[29] Suzuki A, Iwama A, Miyashita H, et al. Role for growth factors and extracellular matrix in controlling differentiation of prospectively isolated hepatic stem cells. Development. 2003; 130(11):2513-2524.

[30] Iwama A, Osawa M, Hirasawa R, et al. Reciprocal roles for CCAAT/enhancer binding protein (C/EBP) and PU.1 transcription factors in Langerhans cell commitment. J Exp Med. 2002;195(5):547-558.