Part I Collaboration with Other iGEM Teams
Overview
In our project, in order to test the function of GCGR predator on a more realistic platform rather than liver carcinoma cell line HepG2, we came up with the idea of mouse primary hepatocytes. After looking up to several paper，adenovirus was finally decided for gene delivery, for its high efficiency in infection of primary cells compared with liposome. However, the safety issue of using adenovirus in our project still is a big thing we need to think over. Considering this matter, we organized some researches about adenovirus
With CSU_CHINA, we discussed about safety and efficiency of methods we have currently used to transport gene circuits into cells and made detailed discussion on how to improve the efficiency and safety of these methods. CSU came up with an idea of transferrin involvement in liposome transfection of breast cancer cells. Transferrin-lipoplexes transfection has been reported to significantly improve transfection efficiency in different cell types such as pancreatic cells, colon cells, etc.[1,2].
This year our team happen to experiment on mouse primary hepatocytes with adenovirus, which is a transferrin enriched cell that, in theory, is suitable for transferrin-liposome mediated transfection. To test such hypothesis and to see if such method can be used on primary cells, we made a collaboration with CSU of testing the transferrin-lipoplex transfection system in this type of cells. If it works well, this method might be a safer way to deliver parts into primary cells.
In summary, We compared transfection efficiency of liposome alone and transferrin-lipoplex, and we also made comparison of transfection efficiency between transferrin-lipoplexes and adenovirus.
Figure 1. CSU_CHINA shared their transferrin with us.
Methods
Results
Figure 2. Fluorescence photography of transfected/infected mouse primary hepatocytes, photographed after . Cell transfected with 2ug of pAdTrack-CMV-GCGR Predator-CMV-GFP, along with 5ul Lipofectamine™2000 + 64ug transferrin/5ul Lipofectamine™2000/64ug transferrin. Infection used 30ul adenovirus with CMV-GCGR Predator-CMV-GFP.
According to current results, transferrin-Lipofectamine™2000 is not able to promote transfection efficiency of Lipofectamine™2000 into mouse primary hepatocytes. Currently for mouse primary hepatocytes, virus like adenovirus is probably still the most efficient way for gene delivery.
[1] Conchita, Tros De Ilarduya, , et al. Antitumoral activity of transferrin-lipoplexes carrying the IL-12 gene in the treatment of colon cancer. Journal of Drug Targeting 14.8, 527-535 (2006):.
[2] Camp, E R , et al. Transferrin Receptor Targeting Nanomedicine Delivering Wild Type P53 Gene Sensitizes Pancreatic Cancer to Gemcitabine Therapy. Cancer gene therapy 179, 2(2013).
Part II Collaboration with ohter organizations
The Second Affiliated Hospital of Chongqing Medical University
We are honored to invite medical workers from the Second Affiliated Hospital of Chongqing medical university to participate in our HP activities. we contacted Guoxiu Li, head nurse of the Department of General Medicine (former Gerontology) of the Second Affiliated Hospital of Chongqing Medical University. With Ms. Li's consent, we conducted a nurse interview and hospital visit.
Simultaneously, several clinicians from the hospital also participated in our interview questionnaire, providing us with valuable advice and pointing out the direction of the project.
Xinqiao Hospital of Army Medical University
Our team had a holistic design of our project but was still searching for proof of our design’s feasibility and looking for aspirations. We decided to turn to experts in both the academic and the medical field. Through the contact of our team member Yiman Deng, we got in touch with experts Dr. Xu Jing and Dr. Zheng Hongting from Xinqiao Hospital. The professores we interviewed highly remark our projects as "open another window to control of blood glucose in a smart and intelligent way " , they also stated the necessity of in vivo testing, and inspired us to use new methods in later experiments.
Red Cross of National University of Defense Technology
One member of our team, Yuhang Ye, is a member of Red Cross of National University of Defense Technology. Through his contact, our team and the Red Cross jointly held the diabetes awareness raising lecture in the old people’s home of Changsha No.1 Social Welfare Institute on June 23rd morning. Our team members handed out self-made brochures to the elders and spread out specific ideas about diabetes and our own project, which also involved the FAQs of living habits of diabetics. Both the Red Cross’s volunteers and the aged drew their attention to diabetes-related common sense.
The Red Cross in Changsha made a decision that they would hold health knowledge dissemination activities with NUDT_CHINA on a regular basis, in this way we hold the belief that our project is beneficial to the community.
