Behavioral pattern separation performance and hippocampal Doublecortin neuron analysis in very old female rats

CHIAVELLINI P; MOREL, GUSTAVO R.; CANATELLI-MALLAT, M; GOYA, RODOLFO G.; LEHMANN M; CLEMENT, J.

Mar del Plata

Congreso; Reunion Anual de Sociedades de Biociencia; 2020

Sociedad Argentina de Investigación Clínica

Cyclic partial cell reprogramming is an emerging avenue of research based on the transient expression of pluripotency genes to generate progressively deeper rejuvenated cells that retain their cell identity. Unlike conventional reprogramming, it can rejuvenate cells ex vivo and in vivo. To this end, we have constructed a regulatable bidirectional adenovector expressing the green fluorescent protein (GFP) and Oct4, Sox2, Klf4 and c-Myc (OSKM) genes. Here, we characterized primary fibroblast cultures derived from young (2 mo.) and old (28 mo.) rats and determined their age-related differences assessing hallmarks as beta-galactosidase, H3K9me3, telomere length, 53BP1, ROS, and H2A. Then, we implemented partial reprogramming by short-term transduction of young and old fibroblasts with our OSKM adenovector and determined whether the above parameters tend to reverse to younger levels in the old cells. Cell aging and identity markers were determined by immunocytochemistry (ICC). Fibroblasts were transduced with our STEMCCA HD adenovector, letting the OSKM genes act for 5 days, and their markers compared with non-treated cells. Cell markers showed the expected age-related alterations. After 5 days of OSKM gene expression, some cell marker levels showed in the old cells a trend to change back to younger levels. Markers of cell identity were not affected by the OSKM treatment. Expression of the pluripotency markers NANOG and SSEA was not detected in the treated cells. Partial reprogramming emerges as a powerful tool for the implementation of in vitro and in vivo rejuvenation keeping cell type identity unchanged.