Cryopreservation of human endothelial cells for vascular tissue engineering

Lehle, Karla and Hoenicka, Markus and Jacobs, V. R. and Schmid, Franz X. and Birnbaum, Dietrich E. (2005) Cryopreservation of human endothelial cells for vascular tissue engineering. CRYOBIOLOGY, 50 (2). pp. 154-161. ISSN 0011-2240,

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Abstract

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% dimethyl sulfoxide in a high-potassium solution, cooled to -80 ° C 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/cm(2) for 3-5 days in order to determine the cell multiplication factor. Fresh and cryopreserved/thawed cells were analyzed for their growth, and their anti-inflammatory and anticoagulant function by using cellular ELISA. Cryopreservation resulted in a retrieval of 66 ± 5% and a viability of 79 ± 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. © 2005 Elsevier Inc. All rights reserved.

Item Type: Article
Uncontrolled Keywords: IN-VITRO ENDOTHELIALIZATION; UMBILICAL-CORD CELLS; ADHESION MOLECULES; PRESERVATION; GRAFTS; EXPRESSION; cryopreservation; dimethyl sulfoxide; endothelial cells; growth; function; tissue engineering
Subjects: 600 Technology > 610 Medical sciences Medicine
Divisions: Medicine > Lehrstuhl für Herz-, Thorax- und herznahe Gefäßchirurgie
Depositing User: Dr. Gernot Deinzer
Date Deposited: 17 May 2021 12:47
Last Modified: 17 May 2021 12:47
URI: https://pred.uni-regensburg.de/id/eprint/36293

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