Optimisation of a screening platform for determining IL-6 inflammatory signalling in the senescence-associated secretory phenotype (SASP)

• DOI: 10.1007/s10522-019-09796-4
• Publication Date: 2019-02-11T05:21:55Z
• Authors: Adam Rolt; Anitha Nair; Lynne S. Cox
• Keywords: Senescence; HEK 293 cells; High-Throughput Screening; Proinflammatory cytokine

Cellular senescence has been shown to be sufficient for the development of multiple age-related pathologies. Senescent cells adopt a secretory phenotype (the SASP) which comprises large number pro-inflammatory cytokines, chemokines and proteases. The SASP itself is thought causative in many pathologies diseases, there growing interest developing seno-modifying agents that can suppress SASP. However, order identify new agents, it necessary conduct moderate high throughput screening with robust assays required outcome. Here, we describe optimisation validation cell-based biosensor HEK cell line measurement IL-6 concentrations within range secreted into conditioned medium by primary senescent fibroblasts, adapted 384 well plate format suitable library applications. We further show assay measure changes secretion dependent on population age, responsive mTOR inhibition cells, reduces SASP, including IL-6. Hence, propose this optimised biosensor, term HEK-SASP, may prove value studies requiring robust, renewable relatively inexpensive measuring factors.