Abstract
Successful establishment of a durable endothelial cell (EC) monolayer inside a ventricular blood sac requires homogeneous coverage of the entire luminal surface with attached cells. For this purpose, a new device was developed that slowly rotates a fully assembled cardiac prosthesis with three degrees of freedom. We seeded ECs derived from human adipose tissue at a density of ≃ 3.5 x 104 cells/cm2 onto the surfaces of polyurethane-made blood sacs and 'ersatz' bladders (consisting of T-25 tissue culture flasks). The kinetics of cell attachment, spreading, and proliferation were determined using video microscopy combined with image analysis and cell viability assays. After 60 min of seeding at 5-10 rotations/hr, the plating efficiency inside the blood sacs was 35.7 ± 11%, with cell viability remaining ≃ 90 ± 5%. After 3 hr, when the plating efficiency reached a plateau (≃ 70%), the rotation was stopped and the ECs were allowed to spread and proliferate under static conditions. Within 48 hr, the entire luminal surface was evenly covered by a confluent EC monolayer. Our long-term studies show that with a proper feeding schedule, such an EC monolayer can be maintained intact in vitro for more than 2 weeks.
Original language | English |
---|---|
Pages (from-to) | M319-M324 |
Journal | ASAIO Journal |
Volume | 40 |
Issue number | 3 |
DOIs | |
State | Published - 1994 |
Keywords
- Adipose Tissue/blood supply
- Biomedical Engineering
- Blood
- Cell Adhesion
- Cell Division
- Cells, Cultured
- Endothelium, Vascular/cytology
- Evaluation Studies as Topic
- Heart-Assist Devices
- Humans
- In Vitro Techniques
- Microscopy, Electron, Scanning
- Rotation