Effects of cryopreservation on cell viability and insulin secretion in a model tissue-engineered pancreatic substitute (TEPS).

Abstract

The use of encapsulated insulin-secreting cells constitutes a promising approach towards the treatment of insulin-dependent diabetes. However, long- term storage for off-the-shelf availability still remains an issue, which can be addressed by cryopreservation. This study investigated cryopreservation of a model tissue-engineered pancreatic substitute by two ice-free cryopreservation (vitrification) solutions (designated VS55 and PEG400) in comparison to a conventional freezing protocol. The model substitute consisted of insulin-secreting mouse insulinoma betaTC3 cells entrapped in calcium alginate/poly-L-lysine/alginate (APA) beads. Cell viability and static insulin secretion from the thawed cryopreserved groups were characterized and compared against fresh controls. Cell viability tests using alamarBlue showed that, compared to the fresh groups, the VS55 had the highest viability (p < 0.05), followed by both the PEG400 (p < 0.001) and the frozen groups (p < 0.001). In response to a square wave of glucose, the static insulin secretion data showed that the VS55 and PEG400 groups had similar induction levels against the fresh group, whereas the frozen group had the poorest secretion rate. Cryosubstitution of capsules showed ice formation in the frozen group but no ice in the vitrified groups. Microscopic observations revealed holes and/or tears within beads subjected to freezing, whereas no such abnormalities were detected in the vitrified samples. Overall, vitrification was found to be a promising preservation procedure for this encapsulated cell system.

• Authors: Neil Mukherjee, Zhenzhen Chen, Athanassios Sambanis, Ying Song
• Journal: Cell transplantation (Cell Transplant) Vol. 14 Issue 7 Pg. 449-56 ( 2005) ISSN: 0963-6897 [Print] United States
• PMID: 16285253 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Biocompatible Materials
• Cryoprotective Agents
• Insulin
• Polyethylene Glycols

Topics

• Animals
• Biocompatible Materials
• Cell Survival (drug effects)
• Cell Transplantation (methods)
• Cryopreservation (methods)
• Cryoprotective Agents (chemistry)
• Insulin (biosynthesis, metabolism)
• Insulin Secretion
• Insulin-Secreting Cells (cytology, drug effects, metabolism)
• Insulinoma
• Mice
• Pancreas, Artificial
• Polyethylene Glycols (chemistry, pharmacology)
• Tissue Engineering