Human umbilical cord cells: a new cell source for cardiovascular tissue engineering

Presented at the Eighth Annual Cardiothoracic Techniques and Technologies Meeting 2002, Miami Beach, FL, Jan 23–26, 2002.
https://doi.org/10.1016/S0003-4975(02)03910-3Get rights and content

Abstract

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 α-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.

Section snippets

Cell isolation and cultivation

Human umbilical cord and saphenous vein sections (2 to 3 cm in length) were washed with Dulbecco’s phosphate buffered saline (Gibco, Rockville, MD) and minced into 1-mm pieces, which 100 × 15-mm Petri dishes (Gibco). Tissue sections were cultured with Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (HyClone) and Gentamycin (Gibco). Medium was replaced at 24 and 72 hours and every 6 days thereafter. Daily progress was monitored by phase-contrast

Histology and immunophenotyping of UCC

Hematoxylin and eosin and Trichrome-masson staining of fixed cells showed elongated cells with fibroblast-like morphology as well as deposition of extracellular matrix throughout the cell culture. Immunohistochemistry revealed intracellular expression of ASMA and vimentin, along with formation of collagen I and III (Fig 1). Cultured UCC did not stain positive for the endothelial cell marker CD 31.

Flow cytometry

Results of the FACS analysis of UCC are shown in Figure 2. The UCC characterization by flow

Comment

Living tissue-engineered constructs appear to be an attractive new concept for cardiovascular surgery, especially for the repair of congenital cardiac defects. The creation of viable substitutes with the ability to regenerate, remodel, and grow would have substantial advantages over currently available materials and prostheses.

In the present study, we evaluated the feasibility of using an alternative autologous cell source—namely, human UCC—for tissue engineering of cardiovascular constructs.

Acknowledgements

The authors thank Manfred Welti, Laboratory for Tissue Engineering and Cell Transplantation, University Hospital Zurich, for his valuable work on cell culture, and Klaus Marquard, Department of Surgical Research, University Hospital Zurich, for providing the scanning electron microscopy pictures. We further thank Dr. Jeroen F Visjager, Swiss Institute of Technology, Zurich, for performing the biomechanical testing. Finally, we are grateful to Annegret Bittermann and Oliver Hoechli, Electron

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