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First published online March 23, 2006
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Submitted on December 16, 2005
Accepted on March 8, 2006

Translational and Clinical Research

CHARACTERISATION AND CLINICAL APPLICATION OF HUMAN CD34+ STEM/PROGENITOR CELL POPULATIONS MOBILISED INTO THE BLOOD BY G-CSF

M.Y. Gordon 1, N. Levicar 2, M. Pai 2, P. Bachellier 3, I. Dimarakis 2, F. Al-Allaf 2, H. M'Hamdi 2, T. Thalji 2, J.P. Welsh 2, S.B. Marley 2, J. Davis 2, F. Dazzi 2, F. Marelli-Berg 2, P. Tait 2, R. Playford 2, L. Jiao 2, S. Jensen 2, J.P. Nicholls 1, A. Ayav 2, M. Nohandani 2, F. Farzaneh 4, J. Gaken 4, R. Dodge 5, M. Alison 6, J.F. Apperley 2, R. Lechler 4, N.A. Habib 1*

1 Faculty of Medicine, Imperial College London, Hammersmith Campus, London, United Kingdom; LiverCyte Limited, London, United Kingdom
2 Faculty of Medicine, Imperial College London, Hammersmith Campus, London, United Kingdom
3 LiverCyte Limited, London, United Kingdom
4 Faculty of Medicine, Kings, Guy's and St Thomas's Medical School, King's College Campus, London, United Kingdom
5 Harvard Medical School, Boston, Massecheusetts; Joslin Diabetes Centre, Las Vegas, Nevada
6 The Royal London Hospital, Diabetes and Metabolic Medicine, London, United Kingdom

* To whom correspondence should be addressed. E-mail: nagy.habib{at}imperial.ac.uk.


  Abstract

A phase I study was performed to determine the safety and tolerability of injecting autologous CD34+ cells into five patients with liver insufficiency. The study was based on the hypothesis that the CD34+ cell population in G-CSF mobilised blood contains a subpopulation of cells with the potential for regenerating damaged tissue. We separated a candidate CD34+ stem cell population from the majority of the CD34+ cells (99%) by adherence to tissue culture plastic. The adherent and non-adherent CD34+ cells were distinct in morphology, immunophenotype and gene expression profile. RT-PCR-based gene expression analysis indicated that the adherent CD34+ cells had the potential to express determinants consistent with liver, pancreas, heart, muscle and nerve cell differentiation as well as hematopoiesis. Overall, the characteristics of the adherent CD34+ cells identify them as a separate putative stem/progenitor cell population. In culture, they produced a population of cells exhibiting diverse morphologies and expressing genes corresponding to multiple tissue types. Encouraged by this evidence that the CD34+ cell population contains cells with the potential to form hepatocyte-like cells, five patients with liver insufficiency were given G-CSF to mobilise their stem cells for collection by leukapheresis. Between 1 x 106 and 2 x 108 CD34+ cells were injected into the portal vein (3 patients) or hepatic artery (2 patients). No complications or specific side effects related to the procedure were observed. Three of the five patients showed improvement in serum bilirubin and four of five in serum albumin. These observations warrant further clinical trials.

Key Words. Stem cells, liver disease, transplantation, regenerative medicine




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