Document Detail


Novel cryopreservation method for dissociated human embryonic stem cells in the presence of a ROCK inhibitor.
MedLine Citation:
PMID:  18716037     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Human embryonic stem cells (hESCs) have potential use in clinical therapy and regenerative medicine. One of the major challenges regarding the application of these cells is the development of an efficient cryopreservation protocol, since current methods, which include slow-freezing-rapid thawing and vitrification of colonies in suspension, present poor viability and high differentiation rates. Dissociated hESC suspensions do not survive cryopreservation because they are susceptible to apoptosis upon cell detachment and dissociation. A selective Rho-associated kinase (ROCK) inhibitor has been reported to increase the survival of dissociated hESCs and their cloning efficiency. METHODS AND RESULTS: Here, we describe a novel method for dissociated hESCs cryopreservation in the presence of the ROCK inhibitor Y-27632. The addition of this inhibitor to the freezing and post-thawing medium significantly increased the survival rate and efficiency of colony formation. Moreover, the hESC colonies obtained after the cryopreservation in the presence of the ROCK inhibitor showed a very low rate of differentiation and a reduced time of recovery. After prolonged culture of frozen-thawed dissociated hESCs, the characteristic properties of pluripotent cells were observed, including normal karyotype, morphological features, marker expression (SSEA-4, TRA-1-60, TRA-1-81 and Oct-4) and the potential to differentiate into derivatives of all three germ layers after embryoid bodies formation. CONCLUSION: This novel method for the cryopreservation of dissociated hESCs may reduce the time required to amplify frozen stocks, and facilitate not only the storage of large numbers of hESCs but also the widespread use of these cells in regenerative medicine.
Authors:
R Martin-Ibañez; C Unger; A Strömberg; D Baker; J M Canals; O Hovatta
Related Documents :
17907947 - Production of viable pigs from fetal somatic stem cells.
18535927 - Controlled genetic modification of stem cells for developing drug discovery tools and n...
20460107 - Clonal expansion of human pluripotent stem cells on gelatin-coated surface.
3396727 - Feeder cells from different sources and conditioned media for recloning of human--mouse...
22802697 - Generation of cd2(+)cd8(+) nk cells from c-kit(+) bone marrow cells in porcine.
10540317 - Microglia induce myelin basic protein-specific t cell anergy or t cell activation, acco...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-08-20
Journal Detail:
Title:  Human reproduction (Oxford, England)     Volume:  23     ISSN:  1460-2350     ISO Abbreviation:  Hum. Reprod.     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2008-11-18     Completed Date:  2009-01-30     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8701199     Medline TA:  Hum Reprod     Country:  England    
Other Details:
Languages:  eng     Pagination:  2744-54     Citation Subset:  IM    
Affiliation:
Departament de Biologia Cellular, Facultat de Medicina, IDIBAPS, Universitat de Barcelona and Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas, Barcelona, Spain.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Amides / pharmacology*
Apoptosis / drug effects
Cell Differentiation / drug effects
Cell Line
Cell Survival / drug effects
Cryopreservation / methods*
Cryoprotective Agents / pharmacology
Embryonic Stem Cells* / drug effects
Humans
Pluripotent Stem Cells / drug effects
Pyridines / pharmacology*
rho-Associated Kinases / antagonists & inhibitors*
Chemical
Reg. No./Substance:
0/Amides; 0/Cryoprotective Agents; 0/Pyridines; 138381-45-0/Y 27632; EC 2.7.11.1/rho-Associated Kinases
Comments/Corrections
Comment In:
Hum Reprod. 2009 Feb;24(2):492; author reply 492-3   [PMID:  19091802 ]

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


Previous Document:  Alpha-adrenoceptor blockade modifies neurally induced atrial arrhythmias.
Next Document:  Fibromuscular differentiation in deeply infiltrating endometriosis is a reaction of resident fibrobl...