In utero xenotransplantation of mice bone marrow-derived stromal/stem cells into fetal rat liver: A preliminary study


Background: Animals can play an important role in preparing tissues for human through the development of xenotransplantation protocols. The most common problem with liver transplantation like any other organ transplantation is organ supply shortage.

Objective: To evaluate the in utero xenotransplantation of mouse bone marrow-derived stromal/stem cells (BMSCs) to the liver of rat fetus to produce mouse liver tissue.

Materials and Methods: BMSCs were isolated and confirmed from enhanced green fluorescent protein (eGFP)-genetic labeled mice. Using a microinjection protocol, mice BMSCs were injected into the liver of rat fetuses in utero on day 14 of pregnancy. After birth, livers were collected and the presence of mice eGFP-positive cells in rat livers was evaluated through polymerase chain reaction.

Results: The eGFP mRNA was detected in the liver of injected infant rats. BMSCs of adult mice were capable to remain functional probably as hepatocyte-like cells in liver of infant rats after in utero xenotransplantation.

Conclusion: BMSCs have the potential for intrauterine xenotransplantation for the treatment of liver dysfunction before birth. This method can also be used for xenoproduction of liver tissue for transplantation.

Key words: Xenotransplantation, Liver, Bone marrow, Stromal/stem cell, Murine.

[1] Zakrzewski W, Dobrzyński M, Szymonowicz M, Rybak Z. Stem cells: past, present, and future. Stem Cell Res Ther 2019; 10: 68–89.

[2] Aliborzi G, Vahdati A, Mehrabani D, Ebrahim Hosseini S, Tamadon A. Isolation, characterization and growth kinetic comparison of bone marrow and adipose tissue mesenchymal stem cells of Guinea pig. Int J Stem Cells 2016; 9: 115–123.

[3] Mehrabani D, Hassanshahi MA, Tamadon A, Zare S, Keshavarz S, Rahmanifar F, et al. Adipose tissuederived mesenchymal stem cells repair germinal cells of seminiferous tubules of busulfan-induced azoospermic rats. J Hum Reprod Sci 2015; 8: 103–110.

[4] Mehrabani D, Rahmanifar F, Mellinejad M, Tamadon A, Dianatpour M, Zare S, et al. Isolation, culture, characterization, and adipogenic differentiation of heifer endometrial mesenchymal stem cells. Comp Clin Pathol 2015; 24: 1159–1164.

[5] Rezaeian L, Hosseini SE, Dianatpour M, Edalatmanesh MA, Tanideh N, Mogheiseh A, et al. Intrauterine xenotransplantation of human Wharton jelly-derived mesenchymal stem cells into the liver of rabbit fetuses: A preliminary study for in vivo expression of the human liver genes. Iran J Basic Med Sci 2018; 21: 89–96.

[6] Mehrabani D, Mahdiyar P, Torabi K, Robati R, Zare S, Dianatpour M, et al. Growth kinetics and characterization of human dental pulp stem cells: Comparison between third molar and first premolar teeth. J Clin Exp Dent 2017; 9: e172–e177.

[7] Mehrabani D, Bahrami Nazarabadi R, Kasraeian M, Tamadon A, Dianatpour M, Vahdati A, et al. Growth kinetics, characterization and plasticity of human menstrual blood stem cells. Iran J Med Sci 2016; 41: 132–139.

[8] Razeghian Jahromi I, Mehrabani D, Mohammadi A, Ghahremani Seno MM, Dianatpour M, Zare S, et al. Emergence of signs of neural cells after exposure of bone marrow-derived mesenchymal stem cells to fetal brain extract. Iran J Basic Med Sci 2017; 20: 301–307.

[9] Rahmanifar F, Tamadon A, Mehrabani D, Zare S, Abasi S, Keshavarz S, et al. Histomorphometric evaluation of treatment of rat azoospermic seminiferous tubules by allotransplantation of bone marrow-derived mesenchymal stem cells. Iran J Basic Med Sci 2016; 19: 653–661.

[10] Khajehahmadi Z, Mehrabani D, Ashraf MJ, Rahmanifar F, Tanideh N, Tamadon A, et al. Healing effect of conditioned media from bone marrow-derived stem cells in thioacetamide-induced liver fibrosis of rat. J Med Sci 2016; 16: 7–15.

[11] Volarevic V, Nurkovic J, Arsenijevic N, Stojkovic M. Concise review: therapeutic potential of mesenchymal stem cells for the treatment of acute liver failure and cirrhosis. Stem Cells 2014; 32: 2818–2823.

[12] Ekser B, Gridelli B, Tector AJ, Cooper DK. Pig liver xenotransplantation as a bridge to allotransplantation: which patients might benefit? Transplantation 2009; 88: 1041–1049.

[13] Muench MO. In utero transplantation: baby steps towards an effective therapy. Bone Marrow Transplant 2005; 35: 537–547.

[14] David AL, Peebles D. Gene therapy for the fetus: is there a future? Best Pract Res Clin Obstet Gynecol 2008; 22: 203–218.

[15] Troeger C, Surbek D, Schoeberlein A, Schatt S, Dudler L, Hahn S, et al. In utero haematopoietic stem cell transplantation. Expriences in mice, sheep and humans. Swiss Med Wkly 2006; 136: 498–503.

[16] Emmert MY, Weber B, Wolint P, Frauenfelder T, Zeisberger SM, Behr L, et al. Intramyocardial transplantation and tracking of human mesenchymal stem cells in a novel intra-uterine pre-immune fetal sheep myocardial infarction model: a proof of concept study. PLoS One 2013; 8: e57759–e57772.

[17] Buse E. Development of the immune system in the cynomolgus monkey: the appropriate model in human targeted toxicology? J Immunotoxicol 2005; 2: 211–216.

[18] Goessling W, Allen RS, Guan X, Jin P, Uchida N, Dovey M, et al. Prostaglandin E2 enhances human cord blood stem cell xenotransplants and shows long-term safety in preclinical nonhuman primate transplant models. Cell Stem Cell 2011; 8: 445–458.

[19] Schultz RD, Dunne HW, Heist CE. Ontogeny of the bovine immune response. Infect Immun 1973; 7: 981–991.

[20] Johnson EH, Al-Habsi KR, Al-Busaidi RR. A review of observations made on select parameters of the camel immune system. Agricultural and Marine Sciences 2013; 18: 1–6.

[21] Abellaneda JM, Ramis G, Martínez-Alarcón L, Majado MJ, Quereda JJ, Herrero-Medrano JM, et al. Generation of human-to-pig chimerism to induce tolerance through transcutaneous in utero injection of cord blood-derived mononuclear cells or human bone marrow mesenchymals cells in a preclinical program of liver xenotransplantation: preliminary results. Transplant Proc 2012; 44: 1574–1578.

[22] Ko JY, Lee J, Lee J, Im GI. Intra-articular xenotransplantation of adipose-derived stromal cells to treat osteoarthritis in a goat model. Tissue Eng Regen Med 2017; 14: 65–71.

[23] Lanning D, Zhu X, Zhai SK, Knight KL. Development of the antibody repertoire in rabbit: gut-associated lymphoid tissue, microbes, and selection. Immunol Rev 2000; 175: 214–228.

[24] Lasso JM, Cano RP, Castro Y, Arenas L, García J, Fernández-Santos ME. Xenotransplantation of human adipose-derived stem cells in the regeneration of a rabbit peripheral nerve. J Plast Reconstr Aesthet Surg 2015; 68: e189–e197.

[25] Pilorz V, Jäckel M, Knudsen K, Trillmich F. The cost of a specific immune response in young guinea pigs. Physiol Behav 2005; 85: 205–211.

[26] Robinson AM, Miller S, Payne N, Boyd R, Sakkal S, Nurgali K. Neuroprotective potential of mesenchymal stem cellbased therapy in acute stages of TNBS-induced colitis in guinea-pigs. PLOS One 2015; 10: e0139023.

[27] Holsapple MP, West LJ, Landreth KS. Species comparison of anatomical and functional immune system development. Birth Defects Res B Dev Reprod Toxicol 2003; 68: 321–334.

[28] Paul A, Srivastava S, Chen G, Shum-Tim D, Prakash S. Functional assessment of adipose stem cells for xenotransplantation using myocardial infarction immunocompetent models: Comparison with bone marrow stem cells. Cell Biochem Biophys 2013; 67: 263–273.

[29] Takeshita K, Motoike S, Kajiya M, Komatsu N, Takewaki M, Ouhara K, et al. Xenotransplantation of interferongamma- pretreated clumps of a human mesenchymal stem cell/extracellular matrix complex induces mouse calvarial bone regeneration. Stem Cell Res Ther 2017; 8: 101–114.

[30] Salehi MS, Jafarzadeh Shirazi MR, Zamiri MJ, Pazhoohi F, Namavar MR, Niazi A, et al. Hypothalamic expression of KiSS1 and RFamide-related peptide-3 mRNAs during the estrous cycle of rats. Int J Fertil Steril 2013; 6: 304–309.

[31] Sabet Sarvestani F, Tamadon A, Koohi-Hosseinabadi O, Nezhad SM, Rahmanifar F, Jafarzadeh Shirazi MR, et al. Expression of RFamide-related peptide-3 (RFRP-3) mRNA in dorsomedial hypothalamic nucleus and KiSS-1 mRNA in arcuate nucleus of rat during pregnancy. Int J Fertil Steril 2014; 8: 333–340.

[32] Bajek A, Olkowska J, Drewa T. Mesenchymal stem cells as a therapeutic tool in tissue and organ regeneration. Postepy Hig Med Dosw 2011; 65: 124–132.

[33] Aurich H, Sgodda M, Kaltwasser P, Vetter M, Weise A, Liehr T, et al. Hepatocyte differentiation of mesenchymal stem cells from human adipose tissue in vitro promotes hepatic integration in vivo. Gut 2009; 58: 570–581.

[34] Ortiz LA, Dutreil M, Fattman C, Pandey AC, Torres G, Go K, et al. Interleukin 1 receptor antagonist mediates the antiinflammatory and antifibrotic effect of mesenchymal stem cells during lung injury. Proc Natl Acad Sci U S A 2007; 104: 11002–11007.

[35] Surbek DV, Holzgreve W, Nicolaides KH. Haematopoietic stem cell transplantation and gene therapy in the fetus: ready for clinical use? Hum Reprod Update 2001; 7: 85– 91.

[36] Mackenzie TC, Shaaban AF, Radu A, Flake AW. Engraftment of bone marrow and fetal liver cells after in utero transplantation in MDX mice. J Pediatr Surg 2002; 37: 1058–1064.

[37] Kang LI, Mars WM, Michalopoulos GK. Signals and cells involved in regulating liver regeneration. Cells 2012; 1: 1261–1292.

[38] Yanger K, Knigin D, Zong Y, Maggs L, Gu G, Akiyama H, et al. Adult hepatocytes are generated by self-duplication rather than stem cell differentiation. Cell Stem Cell 2014; 15: 340–349.

[39] Dutkowski P, Linecker M, DeOliveira ML, Mullhaupt B, Clavien PA. Challenges to liver transplantation and strategies to improve outcomes. Gastroenterology 2015; 148: 307–302.