Systemic administration of a novel human umbilical cord mesenchymal stem cells population accelerates the resolution of acute liver injury.
BACKGROUND
Hepatocytes and stem cells transplantation may be an alternative to liver transplantation in acute or chronic liver disease. We aimed to evaluate the therapeutic potential of mesenchymal stem cells from human umbilical cord (UCMSCs), a readily available source of mesenchymal stem cells, in the CCl4-induced acute liver injury model.


METHODS
Mesenchymal stem cells profile was analyzed by flow cytometry. In order to evaluate the capability of our UCMSCs to differentiate in hepatocytes, cells were seeded on three different supports, untreated plastic support, MatrigelTM and human liver acellular matrix. Cells were analyzed by immunocitochemistry for alpha-fetoprotein and albumin expression, qPCR for hepatocyte markers gene expression, Periodic Acid-Schiff staining for glycogen storage, ELISA for albumin detection and colorimetric assay for urea secretion.To assess the effects of undifferentiated UCMSCs in hepatic regeneration after an acute liver injury, we transplanted them via tail vein in mice injected intraperitoneally with a single dose of CCl4. Livers were analyzed by histological evaluation for damage quantification, immunostaining for Kupffer and stellate cells/liver myofibroblasts activation and for UCMSCs homing. Pro- and anti-inflammatory cytokines gene expression was evaluated by qPCR analysis and antioxidant enzyme activity was measured by catalase quantification.Data were analyzed by Mann-Whitney U-test, Kruskal-Wallis test and Cuzick's test followed by Bonferroni correction for multiple comparisons.


RESULTS
We have standardized the isolation procedure to obtain a cell population with hepatogenic properties prior to in vivo transplantation. When subjected to hepatogenic differentiation on untreated plastic support, UCMSCs differentiated in hepatocyte-like cells as demonstrated by their morphology, progressive up-regulation of mature hepatocyte markers, glycogen storage, albumin and urea secretion. However, cells seeded on 3D-supports showed a minor or negligible differentiation capacity.UCMSCs-transplanted mice showed a more rapid damage resolution, as shown by histological analysis, with a lower inflammation level and an increased catalase activity compared to CCl4-treated mice.


CONCLUSIONS
Our findings show that UCMSCs can be reliably isolated, have hepatogenic properties and following systemic administration are able to accelerate the resolution of an acute liver injury without any differentiation and manipulation. These features make UCMSCs strong candidates for future application in regenerative medicine for human acute liver disease.
Gastroenterology, Department of Surgical, Oncological and Gastroenterological Sciences, Padova University Hospital, Via Giustiniani 2, Padova, 35128, Italy. burra@unipd.it
Patrizia
P
Burra
Diletta
D
Arcidiacono
Debora
D
Bizzaro
Tatiana
T
Chioato
Rosa
R
Di Liddo
Antara
A
Banerjee
Andrea
A
Cappon
Patrizio
P
Bo
Maria Teresa
MT
Conconi
Pier Paolo
PP
Parnigotto
Silvia
S
Mirandola
Enrico
E
Gringeri
Amedeo
A
Carraro
Umberto
U
Cillo
Francesco Paolo
FP
Russo
D055371:Acute Lung Injury / Q000139:chemically induced / Q000473:pathology / Q000601:surgery*; D000418:Albumins / Q000032:analysis; D000818:Animals; D001672:Biocompatible Materials; D015415:Biomarkers / Q000032:analysis; D002251:Carbon Tetrachloride / Q000633:toxicity; D002374:Catalase / Q000032:analysis; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D036101:Cord Blood Stem Cell Transplantation; D016207:Cytokines / Q000096:biosynthesis; D006003:Glycogen / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D008115:Liver Regeneration; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D051379:Mice; D059467:Transcriptome; D014183:Transplantation, Heterologous; D014508:Urea / Q000032:analysis; D000509:alpha-Fetoproteins / Q000032:analysis
D000418:Albumins; D001672:Biocompatible Materials; D015415:Biomarkers; D016207:Cytokines; D000509:alpha-Fetoproteins; D014508:Urea; D006003:Glycogen; D002251:Carbon Tetrachloride; D002374:Catalase