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I has several questions about this protocol. 1. Does the FBS...
YL
YX L May 4, 2021
I has several questions about this protocol. 1. Does the FBS in seeding medium for mouse MHs need to be inactivated before use? 2. Could dexamethasone be removed from the medium since dexamethasone stimulated hepatic differentiation？3. Many other papers included HGF in the culture medium, 
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Takeshi Katsuda Author Answered May 8, 2021
Division of Molecular and Cellular Medicine, National Cancer Center Research Institute, Japan, Japan,
Thank you for your questions. My answers are:
1. Yes, we used heat-inactivated FBS.
2. I have not used medium without dexamethasone, so I cannot answer this question.
3. No, we have not used HGF in our hands.
4. After moving to a new lab in the USA, I also experienced a situation in which mouse hepatocytes grew slower than they used to in my previous lab in Japan. Now, I’m using SHM supplemented with 10%FBS, which seems to improve attachment after plating and proliferation of hepatocytes. I also realized that proliferating small cells are mixture of hepatocyte- and BEC-derived cells. I am writing a paper which clarifies the cellular origin of CLiPs and the phenotypic differences of those cells.
Takeshi Katsuda, PhD (University of Pennsylvania)
1. Yes, we used heat-inactivated FBS.
2. I have not used medium without dexamethasone, so I cannot answer this question.
3. No, we have not used HGF in our hands.
4. After moving to a new lab in the USA, I also experienced a situation in which mouse hepatocytes grew slower than they used to in my previous lab in Japan. Now, I’m using SHM supplemented with 10%FBS, which seems to improve attachment after plating and proliferation of hepatocytes. I also realized that proliferating small cells are mixture of hepatocyte- and BEC-derived cells. I am writing a paper which clarifies the cellular origin of CLiPs and the phenotypic differences of those cells.
Takeshi Katsuda, PhD (University of Pennsylvania)
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