Proceeding of Veterinary and Animal Science Days 2016, 8th- 10th June, Milan, Italy 

HAF © 2013 

Vol. III, No. 1s  ISSN: 2283-3927 

   

 

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Keywords 
Epigenetic, 5-aza-CR, 

Methylation, Plasticity, TET 

genes, Histone 

 

CORRESPONDING AUTHOR 

Elena F.M. Manzoni 

elena.manzoni@unimi.it 

 

JOURNAL HOME PAGE 

riviste.unimi.it/index.php/haf 

5-azacytidine reactivates pluripotency gene 

expression, affects TET2 and histone 

transcription and modifies chromatin 

organization and morphology of mammal 

skin fibroblasts. 

E. F.M. Manzonia* , T. A.L. Brevinib, F. Gandolfia 

aDepartment of Agricultural and Environmental Sciences - Production, 
Landscape, Agroenergy, Università degli Studi di Milano, Via Celoria 2, 20122 
Milan, Italy 

bDepartment of Health, Animal Science and Food Safety, Università degli 
Studi di Milano, Via Celoria 10, 20133 Milan, Italy 

 

 
Abstract 
Phenotype expression is controlled by epigenetic regulations that guide cells through differentiation. 

The process is reversible and cells can be driven back to a higher plasticity state with the use of 

epigenetic modifiers. In this work we exposed skin fibroblasts to the demethylating agent 5-azacytidine 

(5-aza-CR), which is a well-known DNA methyltransferase inhibitor, and has been recently shown to 

increase cell plasticity and facilitate phenotype changes in different cell types (Pennarossa, 2013; Brevini, 

2014; Pennarossa, 2014; Brevini, 2016). Although many aspects controlling its demethylating action have 

been widely investigated, the mechanisms through which 5-aza-CR acts on cell plasticity are still poorly 

understood. At the end of 5-aza-CR treatment, cells were divided in three experimental groups and 

cultured for 24 and 48 hours: A) cells were returned in standard fibroblast medium; B) cells were 

cultured in medium specific for pluripotency maintenance; C) cells were placed in a medium 

encouraging pancreatic differentiation. At the end of the culture period, as expected, we could observe 

a global demethylating effect. In parallel, however we detected a transient upregulation of the 

pluripotency genes OCT4, NANOG and REX1. Increased transcription of TET2 and histones belonging to 

the 1,2,3 and 4 families, together with changes in the expression of enzymes controlling histone 

acetylation were also appreciated. Interestingly, these results were accompanied by morphological and 

ultrastructural changes as well as by chromatin structure modifications. All together our findings 

indicate that 5-aza-CR induced somatic cell transition to a higher plasticity state involves novel cellular 

targets that activate multiple epigenetic regulatory pathways.  

Supported by EFSD and Carraresi Foundation. 
 

References 

Brevini, T. A., Pennarossa, G., Rahman, M.M., Paffoni, A., Antonini, S., Ragni, G., deEguileor, M., Tettamanti, G., Gandolfi,  F., 2014. Morphological and Molecular 

Changes of Human Granulosa Cells Exposed to 5-Azacytidine and Addressed Toward Muscular Differentiation. Stem Cell Rev 10, 633-642. 

Brevini, T. A., Pennarossa, G., Acocella, F., Brizzola, S., Zenobi, A., Gandolfi, F., 2016. Epigenetic conversion of adult dog skin fibroblasts into insulin-secreting cells. 

The Veterinary Journal 211, 52-53. 

Pennarossa, G., Maffei, S., Campagnol, M., Tarantini, L., Gandolfi, F., Brevini, T.A., 2013. Brief demethylation step allows the conversion of adult human skin 

fibroblasts into insulin-secreting cells. Proc Natl Acad Sci U S A 110,  8948-8953. 

Pennarossa, G., Maffei, S., Campagnol, M., Rahman, M.M., Brevini, T.A., Gandolfi, F., 2014. Reprogramming of pig dermal fibroblast into insulin secreting cells by 

a brief exposure to 5-aza-cytidine. Stem Cell Rev 10, 31-43. 

 

 

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