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Nuclear transfer with apoptotic bovine fibroblasts: can programmed cell death be reprogrammed?
MedLine Citation:
PMID:  22686478     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Abstract Cell death by apoptosis is considered to be irreversible. However, reports have indicated that its reversibility is possible if the cells have not yet reached the "point of no return." In order to add new information about this topic, we used cells at different moments of apoptotic process as nuclear donors in somatic cell nuclear transfer (SCNT) in order to test if programmed cell death can be reversed. Adult bovine fibroblasts were treated with 10 μM of staurosporine (STP) for 3 h and analyzed for phosphatidylserine externalization (Annexin assay) and presence of active caspase-9. Annexin-positive (Anx+) and Caspase-9-positive (Casp-9+) cells were isolated by FACS and immediately transferred into enucleated in vitro matured bovine oocytes. After STP treatment, 89.9% of cells were Anx+ (4.6% in control cells; p<0.01) and 24.9% were Casp-9+ (2.4% in control cells; p<0.01). Fusion and cleavage were not affected by the use apoptotic cells (p>0.05). Also, the use of Anx+ cells did not affect blastocyst production compared to control (26.4% vs. 22.9%, respectively; p>0.05). However, blastocyst formation was affected by the use of Casp-9+ cells (12.3%; p<0.05). These findings contribute to the idea of that apoptosis is reversible only at early stages. Additionally, we hypothesize that the "point of no return" for apoptosis may be located around activation of Caspase-9.
Authors:
Moyses Dos Santos Miranda; Fabiana Fernandes Bressan; Tiago Henrique Camara De Bem; Giovana Krempel Fonseca Merighe; Otávio Mitio Ohashi; William Alan King; Flavio Viera Meirelles
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Cellular reprogramming     Volume:  14     ISSN:  2152-4998     ISO Abbreviation:  Cell Reprogram     Publication Date:  2012 Jun 
Date Detail:
Created Date:  2012-06-12     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101528176     Medline TA:  Cell Reprogram     Country:  United States    
Other Details:
Languages:  eng     Pagination:  217-24     Citation Subset:  IM    
Affiliation:
1 Faculdade de Biotecnologia, Universidade Federal do Pará (UFPA), Belém, Pará, Brazil .
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