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Comparison of different cooling rates for fibroblast and keratinocyte cryopreservation
Author(s) -
Naaldijk Yahaira,
FriedrichStöckigt Annett,
Sethe Sebastian,
Stolzing Alexandra
Publication year - 2016
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.1815
Subject(s) - cryopreservation , tissue engineering , fibroblast , suspension culture , biomedical engineering , suspension (topology) , cell culture , andrology , computer science , chemistry , microbiology and biotechnology , biology , engineering , mathematics , embryo , medicine , genetics , homotopy , pure mathematics
Easy, cost‐effective and reliable cryopreservation protocols are crucial for the successful and effective application of tissue engineering. Several different protocols are in use, but no comprehensive comparisons across different machine‐based and manual methods have been made. Here, we compare the effects of different cooling rates on the post‐thaw survival and proliferative capacity of two basic cell lines for skin tissue engineering fibroblasts and keratinocytes, cultured and frozen in suspension or as a monolayer. We demonstrate that effectiveness of cryopreservation cannot be reliably determined immediately after thawing: the results at this stage were not indicative of cell growth in culture 3 days post‐thaw. Cryopreservation of fibroblasts in an adherent state greatly diminishes their subsequent growth potential. This was not observed when freezing in suspension. In keratinocytes, however, adherent freezing is as effective as freezing in suspension, which could lead to significant cost and labour savings in a tissue‐engineering environment. The ‘optimal’ cryopreservation protocol depends on cell type and intended use. Where time, ease and cost are dominant factors, the direct freezing into a nitrogen tank (straight freeze) approach remains a viable method. The most effective solution across the board, as measured by viability 3 days post‐thaw, was the commonly used, freezing container method. Where machine‐controlled cryopreservation is deemed important for tissue‐engineering Good Manufacturing Practice, we present results using a portfolio of different cooling rates, identifying the ‘optimal’ protocol depending on cell type and culture method. Copyright © 2013 John Wiley & Sons, Ltd.