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Cryopreservation of human endothelial cells for vascular tissue engineering.

Author(s) : LEHLE K., HOENICKA M., JACOBS V. R., et al.

Type of article: Article

Summary

To investigate the influence of cryopreservation on endothelial cell growth, morphology, and function human umbilical vein endothelial cells (HUVECs) were frozen following a standard protocol. Cell suspensions were exposed to 10% DMSO in a high-potassium solution, cooled to -80 at 1 °C/min and stored in liquid nitrogen for 7-36 days. Samples were thawed in a 37 °C water bath and the cryoprotectant was removed by serial dilution. The growth of cell suspensions was assayed by culturing 7300 cells/cm2 for 3-5 days to determine the cell multiplication factor. Fresh and cryopreserved/thawed cells were analysed for growth, and anti-inflammatory and anti-coagulant function by using cellular ELISA. Cryopreservation resulted in a retrieval of 66 plus or minus 5% and a viability of 79 plus or minus 3%. Cryopreserved/thawed and fresh cells showed identical doubling times and identical cell counts in the confluent monolayers. However, the lag phase of thawed HUVECs was approximately 36 h longer, resulting in significant differences in the cell multiplication factor at 3 and 5 days after seeding. After expansion to a sufficient cell count the lag phases were identical. Fresh and cryopreserved/thawed cells showed comparable anti-inflammatory and anti-coagulant activity, as judged by the basal and TNF-induced VCAM-1, ICAM-1, E-selectin, and thrombomodulin expression. Cryopreserved/thawed and recultivated endothelial cells are suitable for endothelialization of autologous allograft veins. Such tissue-engineered grafts will offer the necessary clinical safety for those patients who lack autologous material.

Details

  • Original title: Cryopreservation of human endothelial cells for vascular tissue engineering.
  • Record ID : 2006-0464
  • Languages: English
  • Source: Cryobiology - vol. 50 - n. 2
  • Publication date: 2005/04

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