Thank you for visiting my website. The information that populates this site was generated during my graduate school career, and it is my sincere hope that the information will save others attempting to set up a primary mouse hepatocyte system much work and frustration. Primary hepatocytes in general have a reputation as being a challenging system to establish and optimize, and I therefore also hope that this site will make the entire process less daunting and more readily accessible to researchers who have held off pursuing primary mouse hepatocyte work due to the (by no means insignificant) effort required to build the system from scratch.
I started my path towards a doctorate degree during the fall of 2004 at the University of Chicago under the guidance of Dr. Xiao Jian Sun, a gentleman who I consider to this day to be one of the finest scientists I have had the honor of working with. I entered graduate school fresh from college, with ten weeks of laboratory experience and no research background. Our lab studied insulin resistance, and Dr. Sun and I (now, looking back, somewhat naievely) thought that primary mouse hepatocytes would provide an ideal (read: relatively easy), novel means to study the molecular mechanism of insulin resistance in the liver. Unfortunately, we were unable to find any neighboring labs within the University that studied hepatic insulin resistance, or that even had experience working with mouse hepatocytes. We therefore had to resort to the literature for protocols -which, as is almost always the case, did not cater to our specific needs. As we soon found out, there is a rather large gap between isolating cells from the liver and obtaining healthy, happy, and functional hepatocytes.
For brevity's sake, suffice it to say that over the last six years, I have isolated hepatocytes from more mice than I care to remember, and perhaps more importantly, have run a host of functional, as well as molecular assays with said cells to validate the isolation and culture conditions. The table on the front page serves as a partial summary of the readouts I have successfully and consistently run with my hepatocytes.
This web site is a work in progress. I have no illusions about the limits of my knowledge, nor do I claim to have perfected the isolation process. I owe the majority of my success to the pioneers in the field who paved the way for the modern non-recirculating collagenase method of primary hepatocyte isolation; all I can really take credit for is further clarification of particular aspects of the procedure and streamlining certain steps. I cannot even make the claim that I made the information available online- my wonderful, supportive wife, Amanda (the webmaster), deserves all of the credit for that. I therefore welcome all input, critique, and questions, and will do my best to address them.
Finally, I would be remiss if I did not give a special acknowledgement to my current mentor, Dr. Matthew Brady. He was the one who recruited me to the University, and has continued to be a source of unwavering support.
I defended my thesis in February 2011, officially graduated in June 2011, and currently work full-time as a developmental editor in the STM publishing field. While I am no longer generating new data at the bench, I am still very much in touch with basic research, and am committed to continue contributing to science by alternate, although no less useful means.
Sincerely,
Wen Zhang
Contact the scientist at: wz25 [at] uchicago [dot] edu
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