Protocol Protocol for Primary Mouse Hepatocyte Isolation

被引：204

作者：

Charni-Natan, Meital ^{[1
]}

Goldstein, Ido ^{[1
]}

机构：

[1] Hebrew Univ Jerusalem, Inst Biochem Food Sci & Nutr, Robert H Smith Fac Agr Food & Environm, POB 12, IL-7610001 Rehovot, Israel

来源：

STAR PROTOCOLS | 2020年 / 1卷 / 02期

基金：

加拿大健康研究院; 以色列科学基金会;

关键词：

CULTURE; SYSTEMS;

D O I：

10.1016/j.xpro.2020.100086

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Primary hepatocytes are a vital tool in various biomedical research disciplines, serving as an ex vivo model for liver physiology. Obtaining high yields of viable primary mouse hepatocytes is technically challenging, limiting their use. Here, we present an improved protocol based on the classic two-step collagenase perfusion technique. The liver is washed by perfusion, hepatocytes are dissoci-ated by collagenase, separated from other cells, and cultured. This protocol was optimized to significantly reduce procedure duration and improve hepato-cyte yield and viability.

引用

页数：14

共 14 条

[1] Cell-Type-Resolved Quantitative Proteomics of Murine Liver [J].

Azimifar, S. Babak ;

Nagaraj, Nagarjuna ;

Cox, Juergen ;

Mann, Matthias .

CELL METABOLISM, 2014, 20 (06) :1076-1087

[2] Multi-dimensional Transcriptional Remodeling by Physiological Insulin In Vivo [J].

Batista, Thiago M. ;

Garcia-Martin, Ruben ;

Cai, Weikang ;

Konishi, Masahiro ;

O'Neill, Brian T. ;

Sakaguchi, Masaji ;

Kim, Jong Hun ;

Jung, Dae Young ;

Kim, Jason K. ;

Kahn, C. Ronald .

CELL REPORTS, 2019, 26 (12) :3429-+

[3] HIGH-YIELD PREPARATION OF ISOLATED RAT LIVER PARENCHYMAL CELLS - A BIOCHEMICAL AND FINE STRUCTURAL STUDY [J].

BERRY, MN ;

FRIEND, DS .

JOURNAL OF CELL BIOLOGY, 1969, 43 (03) :506-+

[4] Development and utilization of primary hepatocyte culture systems to evaluate metabolism, dna binding, and dna repair of xenobiotics [J].

Casciano, DA .

DRUG METABOLISM REVIEWS, 2000, 32 (01) :1-13

[5] Synergistic gene expression during the acute phase response is characterized by transcription factor assisted loading [J].

Goldstein, Ido ;

Paakinaho, Ville ;

Baek, Songjoon ;

Sung, Myong-Hee ;

Hager, Gordon L. .

NATURE COMMUNICATIONS, 2017, 8

[6] Transcription factor assisted loading and enhancer dynamics dictate the hepatic fasting response [J].

Goldstein, Ido ;

Baek, Songjoon ;

Presman, Diego M. ;

Paakinaho, Ville ;

Swinstead, Erin E. ;

Hager, Gordon L. .

GENOME RESEARCH, 2017, 27 (03) :427-439

[7]

Goldstein I, 2013, CANCER METAB, V1, DOI 10.1186/2049-3002-1-9

[8]

KLAUNIG JE, 1981, IN VITRO CELL DEV B, V17, P913

[9]

Li WC, 2010, METHODS MOL BIOL, V633, P185, DOI 10.1007/978-1-59745-019-5_13

[10] Iron transferrin regulates hepcidin synthesis in primary hepatocyte culture through hemojuvelin and BMP2/4 [J].

Lin, Lan ;

Valore, Erika V. ;

Nemeth, Elizabeta ;

Goodnough, Julia B. ;

Gabatan, Victoria ;

Ganz, Tomas .

BLOOD, 2007, 110 (06) :2182-2189

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