The liver is a very important organ in the human body, which plays the functions of metabolism, detoxification, immunity and so on. Hepatocytes make up about 80-90% of the liver and are the main bearers of liver function. The isolation and culture of primary hepatocytes is one of the most important aspects of mimicking liver diseases and understanding their molecular mechanisms. However, freshly isolated hepatocytes can only be maintained in vitro for about 10-14 days, during which the function of hepatocytes will be continuously lost, and then they will become fibrous mesenchymal cells and eventually apoptosis. This is inconvenient mimicking liver disease.
In 2019, Hongkui Deng's group published a paper "Long-term functional maintenance of primary human hepatocytes in vitro" in the Science, in which they found five compounds that can effectively prolong the time of in vitro culture of primary hepatocytes, and named the combination of these five compounds 5C. These five compounds are Forskolin, SB431542, DAPT, IWP-2 and LDN193189, and their specific information is shown in Table 1, which mainly maintain hepatocytes by affecting related signaling pathways such as differentiation and proliferation. They found that the addition of these 5 compounds to the primary hepatocyte culture process could extend the culture time to 4 weeks and effectively maintain the shape of hepatocytes and the stable expression of related genes.
Primary Human Hepatocytes Maintenance Kit (5C) provides ready-to-use 5C combination reagents, all of which are supplied as stock solutions dissolved in high-purity DMSO. According to the recommended concentration in this manual, this kit can prepare 1 L of culture medium.
|
Compounds
|
CAS Number
|
MW
|
Target
|
|
Forskolin
|
66575-29-9
|
410.50
|
cAMP activator
|
|
SB431542
|
301836-41-9
|
384.39
|
TGF-β/Smad inhibitor
|
|
DAPT
|
208255-80-5
|
432.46
|
Notch inhibitor
|
|
IWP-2
|
686770-61-6
|
466.60
|
Wnt inhibitor
|
|
LDN193189
|
1062368-24-4
|
406.48
|
BMP inhibitor
|
Table 1: Basic information on each compound in the 5C combination
