Document Detail

Construction of autologous human heart valves based on an acellular allograft matrix.
MedLine Citation:
PMID:  12354711     Owner:  NLM     Status:  MEDLINE    
OBJECTIVE: Tissue engineered heart valves based on polymeric or xenogeneic matrices have several disadvantages, such as instability of biodegradable polymeric scaffolds, unknown transfer of animal related infectious diseases, and xenogeneic rejection patterns. To overcome these limitations we developed tissue engineered heart valves based on human matrices reseeded with autologous cells. METHODS AND RESULTS: Aortic (n=5) and pulmonary (n=6) human allografts were harvested from cadavers (6.2+/-3.1 hours after death) under sterile conditions. Homografts stored in Earle's Medium 199 enriched with 100 IU/mL Penicillin-Streptomycin for 2 to 28 days (mean 7.3+/-10.2 days) showed partially preserved cellular viability (MTT assay) and morphological integrity of the extracellular matrix (H-E staining). For decellularization, valves were treated with Trypsin/EDTA resulting in cell-free scaffolds (DNA-assay) with preserved extracellular matrix (confocal microscopy). Primary human venous endothelial cells (HEC) were cultivated and labeled with carboxy-fluorescein diacetate-succinimidyl ester in vitro. After recellularization under fluid conditions, EC were detected on the luminal surfaces of the matrix. They appeared as a monolayer of positively labeled cells for PECAM-1, VE-cadherin and Flk-1. Reseeded EC on the acellular allograft scaffold exhibited high metabolic activity (MTT assay). CONCLUSIONS: Earle's Medium 199 enriched with low concentration of antibiotics represents an excellent medium for long time preservation of extracellular matrix. After complete acellularization with Trypsin/EDTA, recellularization under shear stress conditions of the allogeneic scaffold results in the formation of a viable confluent HEC monolayer. These results represent a promising step toward the construction of autologous heart valves based on acellular human allograft matrix.
Serghei Cebotari; Heike Mertsching; Klaus Kallenbach; Sawa Kostin; Oleg Repin; Aurel Batrinac; Carmen Kleczka; Anatol Ciubotaru; Axel Haverich
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Circulation     Volume:  106     ISSN:  1524-4539     ISO Abbreviation:  Circulation     Publication Date:  2002 Sep 
Date Detail:
Created Date:  2002-09-30     Completed Date:  2002-10-21     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  0147763     Medline TA:  Circulation     Country:  United States    
Other Details:
Languages:  eng     Pagination:  I63-I68     Citation Subset:  AIM; IM    
Leibniz Research Laboratory for Biotechnology and Artificial Organs, Hannover, Germany.
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MeSH Terms
Aortic Valve / anatomy & histology,  chemistry,  metabolism
Cells, Cultured
Collagen Type I / analysis
Endothelium, Vascular / chemistry
Extracellular Matrix / chemistry
Heart Valve Prosthesis*
Microscopy, Fluorescence
Pulmonary Valve / anatomy & histology,  chemistry,  metabolism
Tissue Engineering / methods*
Transplantation, Homologous
Reg. No./Substance:
0/Collagen Type I

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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