Phenotypic characteristics of hybrid cells produced by cell fusion of porcine adrenal chromaffin cells with human mesenchymal stem cells: a preliminary study.
OBJECTIVE
Transplantation of adrenal chromaffin cells (CCs) that release endogenous opioid peptides and catecholamines produces significant antinociceptive effects in patients with terminal cancer pain. In clinical practice, however, obtaining a sufficient number of chromaffin cells may not be possible because of the limited availability of human adrenal tissue. Recent works have shown that fusion of bone marrow-derived cells with differentiated cells can occur spontaneously in vivo and acquire the phenotype of the recipient cells. In this study, we investigated the possibility of producing chromaffin-like cells by fusing human bone marrow-derived mesenchymal stem cells (MeSCs) with post-mitotic porcine CCs in vitro with the application of polyethylene glycol (PEG).


RESULTS
Before cell-to-cell fusion was initiated, MeSCs and CCs were labeled by fluorescence dyes DiO (green) and Dil (red), respectively. The hybrid cells generated by PEG-mediated fusion expressed a DiO and Dil double-staining with estimated fusion efficiency at approximately 40% of the total cell population. Further immunocytochemical examination for tyrosine hydroxylase and methionine enkephalin (markers for CCs) demonstrated positive immuno-reactivity in these hybrid cells 2 weeks post-fusion. More interestingly, some of the hybrid cells showed bromodeoxyuridine (BrdU)-positive immunostaining in the nuclei.


CONCLUSIONS
Our results show that these hybrid cells fused by CCs and MeSCs express some characteristics of the CC phenotype. A subpopulation of these hybrid cells are dividing cells with positive BrdU immunostaining, suggesting that a novel cellular production could be developed by a "reprogramming" mechanism through the application of targeted cell fusion strategies.
Shi
Guangbin
G
The Psychiatric Institute, Department of Psychiatry, College of Medicine, University of Illinois at Chicago, 1601 West Taylor Street, Chicago, IL 60612, USA.
Ma
Ke
K
Pappas
George D
GD
Qu
Tingyu
T
D000313:Adrenal Medulla / Q000166:cytology*; D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation / Q000502:physiology; D002459:Cell Fusion / Q000379:methods; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000187:drug effects / Q000502:physiology*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D006822:Hybrid Cells / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000502:physiology*; D010641:Phenotype; D011092:Polyethylene Glycols / Q000494:pharmacology; D013501:Surface-Active Agents / Q000494:pharmacology; D013552:Swine; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D005456:Fluorescent Dyes; D013501:Surface-Active Agents; D011092:Polyethylene Glycols; D014446:Tyrosine 3-Monooxygenase; D001973:Bromodeoxyuridine