Takahiro Ochiya Division of Molecular and Cellular Medicine, National Cancer Center Research Institute, Japan,

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Yusuke Yamamoto National Cancer Center Research Institute, Tokyo Japan
1 protocol

Marta Prieto-Vila 1 protocol

Kazunori Hosaka Division of Molecular and Cellular Medicine, National Cancer Center Research Institute, Japan, Japan,
1 protocol

Takeshi Katsuda Division of Molecular and Cellular Medicine, National Cancer Center Research Institute, Japan, Japan,
1 protocol

Takahiro Ochiya

Division of Molecular and Cellular Medicine, National Cancer Center Research Institute, Japan, Japan,

Research focus

Cell biology

2 Author merit
Reviewer merit
Editor merit

2 Protocols published

Single-cell qPCR Assay with Massively Parallel Microfluidic System

Authors: Marta Prieto-Vila, Takahiro Ochiya and Yusuke Yamamoto, date: 03/20/2020, view: 3330, Q&A: 0

The single-cell transcriptome is the set of messenger RNA molecules expressed in one cell. It is extremely variable and changes according to external, physical and biochemical conditions. Due to sensitivity shortages, most of genetic studies use ...

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Generation of Chemically Induced Liver Progenitors (CLiPs) from Rat Adult Hepatocytes

Authors: Takeshi Katsuda, Kazunori Hosaka and Takahiro Ochiya, date: 01/20/2018, view: 8823, Q&A: 1

Primary mature hepatocytes (MHs) or their progenitor cells are candidate cell sources for cell transplantation therapy in severe liver diseases. However, stable culture of these cells or generation of equivalent cells from pluripotent stem cells has ...

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YX L

厦门大学

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, have you ever tested it and what is your opinion？4. Only small part of the mouse hepatocytes show a typical progenitor cell morphology after cultured for 8-10 days in our lab, the other cells are large with strange shapes, even though they proliferated , what is your advise？ Thnaks a lot

5/4/2021 7:26:12 PM Reply

Takeshi Katsuda

University of Pennsylvania

Sorry for my late response. I used commercially available collagen-coated plates (IWAKI, Japan), while I prepared matrigel-coated plates by myself.

2022-01-14 08:54:29

YX L

厦门大学

Thank you for your answer，have yor ever tried to coated the plates yourself or you used commercially manufactured pre-coated plates only? I am wondering whether it matters?

2021-05-10 21:15:01

Takeshi Katsuda

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)

2021-05-08 15:25:50