AAGP™ Protects Mouse Embryonic Stem Cells (E14K) During Cryopreservation

Hussein, January 2007

Introduction

Cryopreservation is a technique commonly used to preserve human tissue and cells during long-term storage for their use in transplantation and research. One of the largest problems with stem cell Cryopreservation is maintaining cell number. Anti Aging Glycopeptides (AAGPs™) are small, stable synthetic replicas of antifreeze glycoproteins (AFGPs), a compound found in nature to allow cell survival in fish and insects at freezing temperatures. Thus, the aim of this investigation was to determine the optimal AAGP™ concentration for the protection of mouse embryonic stem cell (E14K) during cryopreservation.

Method

Mouse embryonic stem cells (E14K) were incubated in either 0, 1, 2.5 or 5mg/ml AAGP™ or 4.4mg/ml AAGPs™ (di amino acid) and cryopreserved at -80°C in 20%FBS growth serum containing 10%DMSO at a cell concentration of 600,000 cells/ml. After 2 days of cryopreservation cells were thawed at 37°C. Population cell number was calculated using Trypan blue (4 counts per population). Cell viability/number was calculated as a percentage of cells present prior to cryopreservation (600,000)

Results

Results showed that a concentration of AAGP™ as low as 1mg/ml increased cell viability from 66% (control - no AAGP™ present) to 89%.

Conclusion

This allows us to conclude that concentrations of AAGP™ as low as 1mg/ml can give a 23% increase in cell viability showing that AAGP™, even at low concentrations has cell survival properties during cryopreservation.