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Phenotype Characterization Of Human Di-chimeric Cells For Tolerance Inducing Protocols In Transplantation. Preliminary Study
Joanna Cwykiel MSc, Medhat Askar MD PhD, Maria Siemionow MD PhD DSc
Platic Surgery
2012-02-09
Presenter: Joanna Cwykiel
Affidavit:
New cellular therapy for application in solid organ and vascularized composite allotransplantation will be presented. All of the in vitro assessments were performed by the presenter.
Director Name: Maria Siemionow
Author Category: Attending
Presentation Category: Basic Science Research
Abstract Category: Craniomaxillofacial
How does this presentation meet the established conference educational objectives?
This presentation will address basic science objective of the conference. We will present preclinical evaluation of new cellular therapy that could be applied as a supportive treatment in vascularized composite transplantation.
How will your presentation be used by practicing physicians in the audience?
The knowledge about new supportive cellular therapy for vascularized composite transplantation.
Background: Bone marrow transplantation has already been tested for solid organs and vascularized composite allotransplant (VCA) for modulation of immune responses. We propose a cellular therapy based on ex-vivo created donor-recipient chimeric cells to support VCA and solid organ transplants. The aim was in vitro creation and characterization of phenotype, genotype, and viability of di-chimeric cells (dCC).
Methods: Ten ex vivo fusions of human umbilical cord blood (UCB) cells were performed. Mononuclear cells (MNCs) isolated from UCB originating from 2 different donors were stained separately with PKH26 and PKH67. Fusion was performed using PEG technique. Flow cytometry (FC), (CD3, CD4, CD8, CD19, CD34 and CD90, LIVE/DEAD viability test), confocal microscopy (CM), lymphocytotoxicity (LCT) test for HLA class I antigens, and short tandem repeat (STR) assay were assessed to characterize the phenotype and genotype of dCC.
Results: FC and CM analysis confirmed UCB fusion and creation of human dCC. Using LCT assay we determined that dCC are sharing HLA class I antigens specific for both UCB donors. These results were confirmed by STR, which revealed that dCC were originating (in ~50%) from each UCB donor. Viability assessment showed 99% of cells were viable 3 hours after fusion. Preliminary phenotype characterization showed expression of assessed markers.
Conclusions: We confirmed feasibility of ex vivo fusion of UBC leading to creation of human dCC. We characterized dCC phenotype and viability. This concept of dCC therapy introduces new applications in transplantation. The goal is to induce tolerance in solid organ and VCA transplants.
