An Improved Time- and Labor- Efficient Protocol for Mouse Primary Hepatocyte Isolation.
An Improved Time- and Labor- Efficient Protocol for Mouse Primary Hepatocyte Isolation.
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DOI：
10.3791/61812
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发表时间：
2021-10-25
影响因子：
1.2
通讯作者：
Wu, Sheng
中科院分区：
文献类型：
--
作者：
Feng, Mingxiao;Divall, Sara;Wu, Sheng
原代肝细胞广泛应用于肝脏体外研究，尤其是葡萄糖代谢研究。根据不同的需要，如时间、劳动力、成本和原代肝细胞的使用，对基本技术进行了调整，从而产生了各种原代肝细胞分离方案。然而，原代肝细胞分离的众多步骤和耗时的试剂制备是效率的主要缺点。在比较不同方案的优缺点后，结合各自的优点，制定出快速高效的原代肝细胞分离方案。在大约35分钟内，该方案可以产生与其他方案一样多的健康原代肝细胞，如果不是更多的话。此外，使用分离的原代肝细胞进行的糖代谢实验验证了该方案在体外肝脏代谢研究中的有效性。我们还广泛回顾和分析了本研究中每一步的意义和目的，以便未来的研究人员根据需要进一步优化本方案。
Primary hepatocytes are used extensively in liver in vitro research, especially in glucose metabolism studies. A base technique has been adapted based on different needs, like time, labor, cost, and primary hepatocyte usage, resulting in various primary hepatocyte isolation protocols. However, the numerous steps and time-consuming reagent preparations in primary hepatocyte isolation are major drawbacks for efficiency. After comparing different protocols for their pros and cons, the advantages of each were combined, and a rapid and efficient primary hepatocyte isolation protocol was formulated. Within only ~35 min, this protocol could yield as much, if not more, healthy primary hepatocytes as other protocols. Further, glucose metabolism experiments performed using the isolated primary hepatocytes validated the usefulness of this protocol in in vitro liver metabolism studies. We also extensively reviewed and analyzed the significance and purpose of each step in this study so that future researchers can further optimize this protocol based on needs.
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DOI：
10.3791/56993
发表时间：
2018-02-12
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Cabral F;Miller CM;Kudrna KM;Hass BE;Daubendiek JG;Kellar BM;Harris EN
通讯作者：
Harris EN
DOI：
10.1007/978-1-59745-019-5_13
发表时间：
2010-01-01
期刊：
MOUSE CELL CULTURE: METHODS AND PROTOCOLS
影响因子：
--
作者：
Li, Wan-Chun;Ralphs, Kate L.;Tosh, David
通讯作者：
Tosh, David
影响因子：
6
作者：
Shih, SC;Robinson, GS;Senger, DR
通讯作者：
Senger, DR
影响因子：
1.2
作者：
Korelova, Katerina;Jirouskova, Marketa;Gregor, Martin
通讯作者：
Gregor, Martin
影响因子：
4.7
作者：
Santos, Sofia Duque;Lambertsen, Kate Lykke;Saraiva, Maria Joao
通讯作者：
Saraiva, Maria Joao