International Scholarly Research Notices

Review Article

Cryopreservation-Induced Stress on Long-Term Preserved Articular Cartilage

Table 1

Existing methods for the cryopreservation of tissue engineered constructs.


Preservation method	Storage temperature	Storage medium	Duration of storage	Advantage	Disadvantage	Reference

Domestic freezer	4°C	Culture medium often without any addition of CPA	14–28 days	No ice crystal formation, no CPA addition	Short storage period which is insufficient to carry out immunological and biomechanical assays which are to be carried before transplantation, decrease in cellular activity with an increase in time	Bae et al., 2009 [4] Williams et al., 2003 [5]

Deep freezer	−80°C	CPA or without CPA	30–60 days	No complex device required, low running cost	Critical temperature for preservation of the 3D construct and the sensitive cells seeded on it, uncontrolled freezing, ice crystals formation	Bakhach, 2009 [6]

Controlled rate freezing	A stepwise method can be up to −150°C	CPAs are added	As long as desired when sample is plunged in LN2	Low concentration of CPA, cooling is carried out at a constant cooling rate so as to cause minimum damage to the tissue, allows the cells to dehydrate in equilibrium with the partially frozen cryoprotectants and other extra solutions present in the suspension	Optimized protocols required for each biological product, needs a skilled and experienced person to handle the entire process so that cooling is optimized enough to allow water to efflux out from the cell during extracellular ice formation	Enneking, 1991 [7]

Liquid nitrogen LN2	Ultra low temperature −196°C	CPAs are added	As long as desired with reliable LN2 back up	Rapid cooling method, simple handling, mechanically reliable	Risk of contamination via LN2, LN2 is costly; therefore, a high running cost as regular supply of LN2 is required	Salai et al., 1997 [8], Cetinkaya and Arat, 2011 [9]

Vapour phase LN2	Temperature below −150	CPAs are added	As long as desired with reliable LN2 back up	Eliminate the risk of cross-contamination from other samples and microbes	Vapour phase varies from protocol to protocol and the geometry of the tissue, there is often temperature fluctuation within the vapour phase, volume occupied within the container is nonplanned, regular supply of LN2 is required	Tissue engineering Clemens van blitterswijk—chapter 13 by Lilia Kuleshova and Dietmar Hutmacher [10]