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ctt-journal > Lukashik et al. (Abstract)

Lukashik et al. (Abstract)

Cellular Therapy and Transplantation (CTT), Vol. 3, No. 12
doi: 10.3205/ctt-2011-No12-abstract63

© The Authors. This abstract is provided under the following license: Creative Commons Attribution 3.0 Unported

Abstract accepted for "5th Raisa Gorbacheva Memorial Meeting Hematopoietic Stem Cell Transplantation in Children and Adults", Saint Petersburg, Russia, September 18–20, 2011

Preliminary Program

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Autologous bone marrow mesenchymal stem cell transplantation in patients with liver cirrhosis. A pilot study

Svetlana P. Lukashik, Yanina I. Isaikina, Artur T. Shimanskiy, Oksana N. Romanova, Vladimir M. Cirkunov, Olga V. Aleinikova

Belarusian Research Centre for Pediatric Oncology and Hematology, Minsk, Belarus

Correspondence: Yanina I. Isaikina, Belarusian Research Centre for Pediatric Oncology and Hematology, PO Lesnoe-2, Minsk, 223040, Belarus, E-mail: yaninai@spam is badmail.ru

Abstract

Objective: Morbidity and mortality from cirrhosis is increasing throughout the world. Some pilot clinical studies have demonstrated positive therapeutic results after autologous mesenchymal stem cell (MSCs) infusion in patients with severe liver diseases, but they haven't investigated the morphological changes in the liver after this transplantation.

Aim: To evaluate the potential of MSCs derived from bone marrow to differentiate into hepatocyte-like cells in vitro and to investigate the effect of autologous MSCs transplantation on pathological process in the cirrhosis-affected liver.

Methods: For in vitro investigation, МSCs were isolated from bone marrow samples and cultured in the presence of HGF, bFGF, oncostatin M, ITS, and dexamethasone. After 21 days, morphological assay, real-time polymerase chain reactions and glycogen staining were carried out. After protocol approval by the ethics committee, eight patients (7 with HCV-related cirrhosis, and 1 with autoimmune cirrhosis) with end-stage liver disease were included. Autologous MSCs were taken from bone marrow and expanded for 30–50 days in culture. A median of 115 (24–155) x 106 MSCs were diluted in 5 ml saline and injected under endoscopy into the liver parenchyma at 5 points. Liver function and clinical state were evaluated at baseline and 6 month after MSCs transplantation.

Results: We observed that the morphology of MSCs changed during the cultivation and most of cells had the polygonal form with the central nucleus similar to hepatocytes on the 21st day. Hepatocyte-like cells showed positive staining on glycogen, which differs from MSCs. The analysis of an expression of hepatocyte-specific markers revealed the increase of HNF1α, CYP3α4, and albumin expression levels (p<0.05) and an insignificant growth of CK18 expression on the 21st day of differentiation.

One patient was withdrawn from the study 4 months after MSCs transplantation due to tuberculosis. The general condition of all patients improved in a month after MSCs transplantation. Six months after MSCs transplantation the improvement of functional liver tests was observed: serum ALT decreased from 120.71±25.69 to 91.66±17.83 U/l, and bilirubin decreased from 32.18±5.51 µmol/l to 26.71±6.14 µmol/l (p<0.05). Morphological changes were characterized by suppression of fibrogenesis (reduction of sinusoids “capillarization” and quantity of myofibroblasts). Electron-microscope data and PCNA evidenced positive proliferative-reparative processes in hepatocytes (the regeneration of cellular organelles structure) and “heterogeneity” of hepatocytes in the liver tissue. There were no adverse effects in any patient.

Conclusion: These in vitro studies show that МSCs can undergo the process of differentiation into hepatocyte-like cells, which have the morphology of hepatocytes, express various hepatocyte-specific markers, and have hepatocyte-specific bioactivities. Therapeutic use of intraparenchymal injection of MSCs is safe and feasible and may improve liver function and reparative processes in hepatocytes in patients with liver cirrhosis. We assume the intraparenchymal method promotes an increase of MSCs homing in certain sites of the liver, and objective interpreting of results of liver biopsy material studies before and after MSCs injection.

Keywords: mesenchymal stem cells, cirrhosis, transplantation