In utero transplantation of human hematopoietic stem/progenitor cells partially repairs injured liver in mice

Abstract

Abstract. The aim of this study is to establish a novel mouse model with high achievement and chimerism by in utero transplantation of human hematopoietic stem/progenitor cells and to explore the possibility that human adult hematopoietic stem/progenitor cells can differentiate into hepatocyte-like cells and partially repair the liver damage induced by carbon tetrachloride (CCl 4 ). Mononuclear cells (MNCs) were isolated from fresh human umbilical cord blood (hUCB) and CD34 + cells were enriched from the MNCs by magnetic cell isolation. These cells were injected respectively into the fetal mice at 11-13 days of gestation. At one month after birth, the specific markers of human cells, human ·-satellite sequence (h17·), CD14, CD34, CD45, and GPA were detected by PCR and FACS. At three and six months after birth, the established human-mouse chimeras were administered with CCl 4 by intraperitoneal injection. The biochemical markers (ALT, AST, ALP, albumin) in serum were determined and human hepatocyte-specific proteins, such as human albumin, hepatocyte nuclear factor-4, hepatocyte specific antigen, tryptophan 2,3-dioxygenase and · fetoprotein were analyzed by PCR, RT-PCR, real-time PCR and immunohistochemistry staining, respectively. More than 77% of recipients demonstrated human-mouse chimera. Significantly, hUCB hematopoietic stem/progenitor cells may differentiate into human hepatocytelike cells with evidence of the expression of human hepatocytespecific proteins as well as partially repair or protect liver damage induced by CCl 4 . The mouse model described in this article provides a useful tool for the studies of regeneration of human hepatocyte-like cells from adult hematopoietic stem/ progenitor cells as well as facilitates the therapeutic potential for liver diseases or damage by in utero transplantation.

Phenotypic changes of human cells in human-rat liver during partial hepatectomy-induced regeneration

“…In contrast to the immunodeficient mice harboring a humanized liver, HAC in liver humanization possess and potential human immune system can be reconstituted with donor-derived human differentiated cells (data not shown). Besides the functional donor-derived human mature HLCs positive for human Alb, AFP, HNF-4, etc, throughout the host liver parenchyma, an "ideal humananimal chimeric liver" can also synthesize and secrete human Alb, ALT, AST and ALP into the circulation of host animals [2,6,[18][19][20]22,24] . Our findings demonstrate that donor-derived human mature hepatocytes, endothelial and biliary duct cells in human-rat chimeric liver, constitute the "humanized hepatic functional units" in animal liver [2,19,20] .…”

“…In utero or blastocyst transplantation of hSCs from multiple sources can form all possible donor-derived human cell types in the recipient liver, which in turn constitute human hepatic parenchymal and stromal components [2,6,[18][19][20]22,24] . In contrast to the immunodeficient mice harboring a humanized liver, HAC in liver humanization possess and potential human immune system can be reconstituted with donor-derived human differentiated cells (data not shown).…”

“…Moreover, such a "HAC liver" can also produce and secrete human Alb, alanine aminotransferase (ALT), aspartic acid aminotransferase (AST) and alkaline phosphatase (ALP) in the circulation of host mice [18] and sheep [2,6] that have undergone transplantation.…”

“…In utero transplantation of hSCs, such as human hematopoietic stem cells and mesenchymal stem cells, into fetal sheep [2,6] , goats [24] , rats [19,20] , and mice [18,22] or blastocyst transplantation of hSCs into mice [22] , has led to the establishment of non-injury human-animal xenograft models carrying humanized liver, in which a significant number of functional donor-derived human mature hepatocyte-like cells (HLCs) stained positively for human albumin (Alb), alpha fetoprotein (AFP) and hepatocyte nuclear factor-4 can be found. Moreover, such a "HAC liver" can also produce and secrete human Alb, alanine aminotransferase (ALT), aspartic acid aminotransferase (AST) and alkaline phosphatase (ALP) in the circulation of host mice [18] and sheep [2,6] that have undergone transplantation.…”

Cell therapy for the diseased liver: from stem cell biology to novel models for hepatotropic human pathogens

“…A particularly interesting set of experiments involved intra-fetal injections of human UCB cells into mice, rats, sheep and goats (Almeida-Porada et al, 2004;Turrini et al, 2005;Zeng et al, 2005;Qian et al, 2006;Zeng et al, 2006;Sun et al, 2007) (Table 4). Cell engraftment into fetal liver would be expected to preclude the necessity of suppression of the immune response and of induced liver damage to obtain an environment appropriate for cell proliferation.…”