Reevaluation of bone marrow-derived cells as a source for hepatocyte regeneration.
We have investigated the contribution of intrasplenic bone marrow transplants or in vivo mobilized hematopoietic stem cells to the formation of hepatocytes in normal and injured liver. Direct intrasplenic injections of bone marrow mononuclear cells (5 x 10(5) cells), Scal+/lin- hematopoietic stem cells (5 x 10(3)) cells, and highly purified "side population" hematopoietic stem cells (5 x 10(3)) derived from enhanced green fluorescent protein (EGFP)-transgenic mice [C57Bl/6-TgN(ActbEGFP)1Osb] were performed in normal C57Bl/6 mice (n = 6) and in C57Bl/6 mice following two thirds hepatectomy (n = 8). To test the effect of mobilized stem cells on transdifferentiation, C57Bl/6 mice (n = 12) were lethally irradiated and reconstituted with EGFP-positive bone marrow mononuclear cells in a second series of experiments. Eight to 12 weeks after bone marrow transplantation a subset of mice (n = 3 in each group) received either rhG-CSF for hematopoietic stem cell mobilization, rhG-CSF combined with an intraperitoneal application of carbon tetrachloride (CCl4) as hepatocyte regeneration stimulus, or CCl4 alone. All mice were completely perfused with PBS to remove circulating nonorgan cells for analyses 4 weeks later. Liver as well as heart, intestine, spleen, and kidney tissue was analyzed for the presence of EGFP-transgenic cells. In 100 sections (2.3 x 10(7) cells) of any recipient mouse no EGFP-positive hepatocytes were detected either by analysis of native EGFP fluorescence or by immunofluorescence analysis with anti-EGFP and antidipeptidyl peptidase (DPP) IV antibodies. EGFP-transgenic cells resembling heart, kidney, or intestinal cells could also not be proven. The results demonstrate that there is little or no contribution of bone marrow-derived cells to the regeneration of normal and injured liver in the animal models used. Thus, potential therapeutic prospects of hematopoietic stem cell therapy for liver disease have to be critically reassessed.
Cantz
Tobias
T
Laboratory of Cell and Gene Therapy, Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
Sharma
Amar Deep
AD
Jochheim-Richter
Andrea
A
Arseniev
Lubomir
L
Klein
Christoph
C
Manns
Michael P
MP
Ott
Michael
M
D000818:Animals; D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D016026:Bone Marrow Transplantation; D002251:Carbon Tetrachloride / Q000494:pharmacology; D056486:Chemical and Drug Induced Liver Injury; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000032:analysis; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology; D006498:Hepatectomy; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects; D007272:Injections, Intralymphatic; D008107:Liver Diseases / Q000628:therapy*; D008115:Liver Regeneration / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D011994:Recombinant Proteins; D013154:Spleen / Q000166:cytology; D014916:Whole-Body Irradiation
D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D016179:Granulocyte Colony-Stimulating Factor; D049452:Green Fluorescent Proteins; D002251:Carbon Tetrachloride; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases