Human umbilical cord cells: a new cell source for cardiovascular tissue engineering.
BACKGROUND
Tissue engineering of viable, autologous cardiovascular constructs with the potential to grow, repair, and remodel represents a promising new concept for cardiac surgery, especially for pediatric patients. Currently, vascular myofibroblast cells (VC) represent an established cell source for cardiovascular tissue engineering. Cell isolation requires the invasive harvesting of venous or arterial vessel segments before scaffold seeding, a technique that may not be preferable, particularly in pediatric patients. In this study, we investigated the feasibility of using umbilical cord cells (UCC) as an alternative autologous cell source for cardiovascular tissue engineering.


METHODS
Human UCC were isolated from umbilical cord segments and expanded in culture. The cells were sequentially seeded on bioabsorbable copolymer patches (n = 5) and grown in vitro in laminar flow for 14 days. The UCC were characterized by flow cytometry (FACS), histology, immunohistochemistry, and proliferation assays and were compared to saphenous vein-derived VC. Morphologic analysis of the UCC-seeded copolymer patches included histology and both transmission and scanning electron microscopy. Characterization of the extracellular matrix was performed by immunohistochemistry and quantitative extracellular matrix protein assays. The tissue-engineered UCC patches were biomechanically evaluated using uniaxial stress testing and were compared to native tissue.


RESULTS
We found that isolated UCC show a fibroblast-like morphology and superior cell growth compared to VC. Phenotype analysis revealed positive signals for alpha-smooth muscle actin (ASMA), desmin, and vimentin. Histology and immunohistochemistry of seeded polymers showed layered tissue formation containing collagen I, III, and glycoaminoglycans. Transmission electron microscopy showed viable myofibroblasts and the deposition of collagen fibrils. A confluent tissue surface was observed during scanning electron microscopy. Glycoaminoglycan content did not reach values of native tissue, whereas cell content was increased. The biomechanical properties of the tissue-engineered constructs approached native tissue values.


CONCLUSIONS
Tissue engineering of cardiovascular constructs using UCC is feasible in an in vitro environment. The UCC demonstrated excellent growth properties and tissue formation with mechanical properties approaching native tissue. It appears that UCC represent a promising alternative autologous cell source for cardiovascular tissue engineering, offering the additional benefits of using juvenile cells and avoiding the invasive harvesting of intact vascular structures.
Kadner
Alexander
A
Clinic for Cardiovascular Surgery, Department of Gynecology and Obstetrics, University Hospital, Zurich, Switzerland. alexander.kadner@chi.usz.ch
Hoerstrup
Simon P
SP
Tracy
Jay
J
Breymann
Christian
C
Maurus
Christine F
CF
Melnitchouk
Serguei
S
Kadner
Gregor
G
Zund
Gregor
G
Turina
Marko
M
D000199:Actins / Q000032:analysis; D001696:Biomechanical Phenomena; D002319:Cardiovascular System; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003094:Collagen / Q000032:analysis; D003893:Desmin / Q000032:analysis; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D023822:Tissue Engineering / Q000379:methods*; D014746:Vimentin / Q000032:analysis
D000199:Actins; D003893:Desmin; D016326:Extracellular Matrix Proteins; D014746:Vimentin; D003094:Collagen