Document Detail


Vitrification by ultra-fast cooling at a low concentration of cryoprotectants in a quartz micro-capillary: a study using murine embryonic stem cells.
MedLine Citation:
PMID:  18462712     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Conventional cryopreservation protocols for slow-freezing or vitrification involve cell injury due to ice formation/cell dehydration or toxicity of high cryoprotectant (CPA) concentrations, respectively. In this study, we developed a novel cryopreservation technique to achieve ultra-fast cooling rates using a quartz micro-capillary (QMC). The QMC enabled vitrification of murine embryonic stem (ES) cells using an intracellular cryoprotectant concentration in the range used for slowing freezing (1-2M). The cryoprotectants used included 2M 1,2-propanediol (PROH, cell membrane permeable) and 0.5M extracellular trehalose (cell membrane impermeable). More than 70% of the murine ES cells post-vitrification attached with respect to non-frozen control cells, and the proliferation rates of the two groups were similar. Preservation of undifferentiated properties of the pluripotent murine ES cells post-vitrification cryopreservation was verified using three different types of assays: the expression of transcription factor Oct-4, the presentation of the membrane surface glycoprotein SSEA-1, and the elevated expression of the intracellular enzyme alkaline phosphatase. These results indicate that vitrification at a low concentration (2M) of intracellular cryoprotectants is a viable and effective approach for the cryopreservation of murine embryonic stem cells.
Authors:
Xiaoming He; Eric Y H Park; Alex Fowler; Martin L Yarmush; Mehmet Toner
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2008-03-30
Journal Detail:
Title:  Cryobiology     Volume:  56     ISSN:  1090-2392     ISO Abbreviation:  Cryobiology     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-05-21     Completed Date:  2008-07-01     Revised Date:  2013-06-05    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  223-32     Citation Subset:  IM    
Affiliation:
Center for Engineering in Medicine and Department of Surgical Service, Massachusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, 51 Blossom Street, Boston, MA 02114, USA. xmhe@sc.edu
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Alkaline Phosphatase / analysis
Animals
Antigens, CD15 / analysis
Cell Adhesion
Cell Differentiation
Cell Proliferation
Cell Survival
Cells, Cultured
Cryopreservation / methods*
Cryoprotective Agents / pharmacology*
Embryonic Stem Cells / cytology*
Ethylene Glycol / pharmacology
Mice
Models, Theoretical
Octamer Transcription Factor-3 / analysis
Plastics / chemistry
Propylene Glycol / pharmacology
Quartz / chemistry*
Thermal Conductivity
Trehalose / pharmacology
Grant Support
ID/Acronym/Agency:
EB002340/EB/NIBIB NIH HHS; R01 EB002340-05/EB/NIBIB NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, CD15; 0/Cryoprotective Agents; 0/Octamer Transcription Factor-3; 0/Plastics; 107-21-1/Ethylene Glycol; 14808-60-7/Quartz; 57-55-6/Propylene Glycol; 99-20-7/Trehalose; EC 3.1.3.1/Alkaline Phosphatase
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Incremental HMM training applied to ECG signal analysis.
Next Document:  Loss of unc45a precipitates arteriovenous shunting in the aortic arches.