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  • Manipulating and tracking single hepatocyte behavior during mouse liver regeneration by performing hydrodynamic tail vein injection.

Manipulating and tracking single hepatocyte behavior during mouse liver regeneration by performing hydrodynamic tail vein injection.

STAR protocols (2021-04-27)
Shuang Wang, Marina Ruiz de Galarreta, Kirsten C Sadler, Amaia Lujambio
RÉSUMÉ

Studies to identify genes relevant to mammalian hepatocyte biology in vivo are largely carried out using germline genetic-engineering approaches, which can be costly and time-consuming. We describe hydrodynamic tail vein injection as an alternative approach to introduce genetic elements into hepatocytes. Transfected hepatocytes can then be traced with a GFP reporter enabling the use of immunohistochemistry and FACS sorting to examine the changes in hepatocyte gene expression and proliferation during liver regeneration induced by 2/3 partial hepatectomy (PH). For complete details on the use and execution of this protocol, please refer to Wang et al. (2019).

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Solution de formol, tamponnée à pH neutre (10 %), histological tissue fixative
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DAPI, for nucleic acid staining
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Albumine de sérum bovin, lyophilized powder, crystallized, ≥98.0% (GE)
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SOC Medium, For use in transformation
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Ampicilline sodium salt
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Iodure de propidium solution
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LB Broth with agar (Lennox), EZMix powder microbial growth medium