TY - JOUR
T1 - Transient alteration of gene expression in adipose-derived stem cells using liposomal-driven protein extracts
AU - Campillo, Noelia
AU - Arribas, María I.
AU - Vicente-Salar, Nestor
AU - Catania, Angela
AU - Ramírez-Domínguez, Miriam
AU - Reig, Juan A.
AU - Domínguez-Bendala, Juan
AU - Micol, Vicente
AU - Roche, Enrique
N1 - Funding Information:
Enrique Roche is recipient of the following grants: Instituto de Salud Carlos III-FEDER (PS09/01093) and Fundacion Salud 2000-Merck Serono. Vicente Micol and Enrique Roche are recipients of a grant PROME-TEO/2012/007 from Generalitat Valenciana and are members of the ‘‘Centro de Investigación Biomédica en Red de Fisiopatología de la Obesidad y Nutrición’’ CIBERobn (CB12/03/30038). Maria I Arribas is recipient of a fellowship of the VALi+d Program from Generalitat Valenciana (APOSTD/2012/021). Miriam Ramírez-Domínguez is member of the Spanish Biomedical Research Centre in Diabetes and Associated Metabolic Disorders (CIBERDEM). Authors declare no conflicts of interest. We want to thank to Dr. Jose Villalain for the use of the LiposoFast®.
PY - 2014/3
Y1 - 2014/3
N2 - Recent works have proposed the use of proteins instead of DNA/RNA as molecular tools for the modulation of gene expression patterns in stem cells. To obtain insulin-producing cells, certain proteins including Pdx-1, Isl-1, Pax-6, and Neuro-D, can be expressed or modified with transduction domains in order to penetrate and reprogram target cells. However, numerous aspects must be taken in account, including the fact that not all proteins may be used in transduction protocols. Thus, new methods must be developed. In this report we studied whether liposomes can work as carriers to introduce reprogramming protein extracts inside adipose tissue mesenchymal stem cells. To this end, the cells were incubated for 8 h in the presence of liposomes containing MIN-6 protein extracts. Charged liposomes fused with 70% of the cells, transferring their contents and transiently (up to 48 h) changing the gene expression pattern for specific endocrine pancreatic genes coding for Insulin-I, Insulin-II, Glucagon, Isl-1, MafA, and Nkx-6.1. No expression of Foxa-2, Pax-6, and Pdx-1 was observed. Although preliminary, the obtained results suggest that the use of liposomes for reprogramming techniques opens new possibilities for cell therapy.
AB - Recent works have proposed the use of proteins instead of DNA/RNA as molecular tools for the modulation of gene expression patterns in stem cells. To obtain insulin-producing cells, certain proteins including Pdx-1, Isl-1, Pax-6, and Neuro-D, can be expressed or modified with transduction domains in order to penetrate and reprogram target cells. However, numerous aspects must be taken in account, including the fact that not all proteins may be used in transduction protocols. Thus, new methods must be developed. In this report we studied whether liposomes can work as carriers to introduce reprogramming protein extracts inside adipose tissue mesenchymal stem cells. To this end, the cells were incubated for 8 h in the presence of liposomes containing MIN-6 protein extracts. Charged liposomes fused with 70% of the cells, transferring their contents and transiently (up to 48 h) changing the gene expression pattern for specific endocrine pancreatic genes coding for Insulin-I, Insulin-II, Glucagon, Isl-1, MafA, and Nkx-6.1. No expression of Foxa-2, Pax-6, and Pdx-1 was observed. Although preliminary, the obtained results suggest that the use of liposomes for reprogramming techniques opens new possibilities for cell therapy.
KW - Adipose tissue
KW - Adult stem cells
KW - Cell reprogramming
KW - Liposomes
KW - Protein transduction
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U2 - 10.1007/s12195-013-0298-3
DO - 10.1007/s12195-013-0298-3
M3 - Article
AN - SCOPUS:84899490211
VL - 7
SP - 145
EP - 154
JO - Cellular and Molecular Bioengineering
JF - Cellular and Molecular Bioengineering
SN - 1865-5025
IS - 1
ER -