Senescence-induced immunophenotype, gene expression and electrophysiology changes in human amniocytes.

Senescence-induced immunophenotype, gene expression and electrophysiology changes in human amniocytes. J Cell Mol Med. 2019 Sep 03;: Authors: Airini R, Iordache F, Alexandru D, Savu L, Epureanu FB, Mihailescu D, Amuzescu B, Maniu H Abstract The aim of the study was to evidence replicative senescence-induced changes in human amniocytes via flow cytometry, quantitative reverse-transcription-polymerase chain reaction (qRT-PCR) and automated/manual patch-clamp. Both cryopreserved and senescent amniocytes cultured in BIO-AMF-2 medium featured high percentages of pluripotency cell surface antigens SSEA-1, SSEA-4, TRA1-60, TRA1-81 (assessed by flow cytometry) and expression of pluripotency markers Oct4 (Pou5f1) and Nanog (by qRT-PCR). We demonstrated in senescent vs cryopreserved amniocytes decreases in mesenchymal stem cell surface markers. Senescence-associated β-galactosidase stained only senescent amniocytes, and they showed no deoxyuridine incorporation. The gene expression profile revealed a secretory phenotype of senescent amniocytes (increased interleukin (IL)-1α, IL-6, IL-8, transforming growth factor β, nuclear factor κB p65 expression), increases for cell cycle-regulating genes (p16INK4A ), cytoskeletal elements (β-actin); HMGB1, c-Myc, Bcl-2 showed reduced changes and p21, MDM2 decreased. Via patch-clamp we identified five ion current components: outward rectifier K+ current, an inactivatable component, big conductance Ca2+...
Source: J Cell Mol Med - Category: Molecular Biology Authors: Tags: J Cell Mol Med Source Type: research