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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
ABSTRAKT

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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SOC Medium, For use in transformation
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Ampicillin sodium salt
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DAPI, for nucleic acid staining
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Propidium iodide solution
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Formalin solution, neutral buffered, 10%, histological tissue fixative
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LB Broth with agar (Lennox), EZMix powder microbial growth medium
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Bovine Serum Albumin, lyophilized powder, crystallized, ≥98.0% (GE)