"Universal" vitrification of cells by ultra-fast cooling.
Long-term preservation of live cells is critical for a broad range of clinical and research applications. With the increasing diversity of cells that need to be preserved (e.g. oocytes, stem and other primary cells, genetically modified cells), careful optimization of preservation protocols becomes tedious and poses significant limitations for all but the most expert users. To address the challenge of long-term storage of critical, heterogeneous cell types, we propose a universal protocol for cell vitrification that is independent of cell phenotype and uses only low concentrations of cryoprotectant (1.5 M PROH and 0.5 M trehalose). We employed industrial grade microcapillaries made of highly conductive fused silica, which are commonly used for analytical chemistry applications. The minimal mass and thermal inertia of the microcapillaries enabled us to achieve ultrafast cooling rates up to 4,000 K/s. Using the same low, non-toxic concentration of cryoprotectant, we demonstrate high recovery and viability rates after vitrification for human mammary epithelial cells, rat hepatocytes, tumor cells from pleural effusions, and multiple cancer cell lines.
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA.
Yun Seok
YS
Heo
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA.
Sunitha
S
Nagrath
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA.
Alessandra L
AL
Moore
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA.
Mahnaz
M
Zeinali
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA.
Daniel
D
Irimia
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA ; Massachusetts General Hospital, Cancer Center, Harvard Medical School, Boston, MA.
Shannon L
SL
Stott
Massachusetts General Hospital, Obstetrics and Gynecology Services, Harvard Medical School, Boston, MA.
Thomas L
TL
Toth
BioMEMS Resource Center, Mass achusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, MA.
Mehmet
M
Toner