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2.2 Laboratory-scale bioreactor cell culture At the laboratory scale, MRC-5 cells were cultured in 3 L glass bioreactors with a BioFlo 320 controller (Cat#: 1379962911, Eppendorf, Hamburg, Germany) on 1 g/L Cytodex 1 microcarriers (Cat#: 17548702, Cytiva, Marlborough, MA) or in iCELLis Nano fixed-bed bioreactors (Pall Corporation, Port Washington, NY) with a 10 cm bed height with WMEM and 10% BCS controlled at a target temperature of 37C and pH of 7.2 as described previously ( 250 HT reactors (Sartorius AG, Gttingen, Germany) with 1 g/L Cytodex 1 microcarriers were operated for oncolytic virus production similar to a published method ( 250 HT bioreactors underwent an 80% medium exchange into WMEM without serum for a final concentration of 2% (v/v) BCS

Immunofluorescence staining of senescent cells Normal and DOX-induced senescent ATDC5 cells were fixed with 4% paraformaldehyde for 15 min, permeabilized with phosphate-buffered saline (PBS) containing 0.25% Triton X-100 for 10 min, and then blocked with 4% (v/v) BSA for 30 min at RT

These lifestyle predispositions can create an environment that allows for certain root cause factors to develop, which begins the self-perpetuating cycle that is Hashimotos

Formal analysis: Zhuang Zhou

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