Primary Culture Of Lung Adenocarcinoma Cells And Different Oncogenicity In Vitro Of Lung Adenocarcinoma Cells

Objectives To find a high productive method in lung adenocarcinoma primary culture in vitro,we compare two different methods in cell seperation and two culture mediums.To prove the lung adenocarcinoma cell's oncogenic capability in vitro.Methods Five lung adenocarcinoma samples were obtained,then divided randomly into four parts.Each one was dealed with the following protocol respectively:A:explant cell culture+serum medium;B: mechanical method+serum medium;C:explant cell culture+serum-free medium;D:mechanical method+serum-free medium.Observe the growth situation under the phase contrast microscope.Count the total cellular score and the score of lung adenocarcinoma cell in different time points,then calculate the percentage of the lung adenocarcinoma cell. Analysis the date by factor analysis of two factors.Clone the lung adenocarcinoma cells which were get from our culture in agar,count the number of cell cluster which has more than 50 cells,calculate the cloning efficiency.Results There are more cells obtained by mechanical method (49.41±2.7×10~4)than explant cell culture(41.72±5.1×10~4),but no stastical significance(P=0.137).Lung adenocarcinoma cells have a higher proportion with mechanical method(59.49±4.0%)than with explant cell culture(37.67±3.1%)(P=0.021).Serum culture has a more promotive effect in cell proliferation than serum-free medium(P=0.035) Serum-free medium has an obvious effect on inhibition to fibroblast (P=0.005).The cultured lung adenocarcinoma cells cloning efficiency is 20%.Conclusion Mechanical method and the Serum-free medium have advantage in primary cell culture of lung adenocarcinoma.The cultured lung adenocarcinoma cells have oncogenic capability in vitro,The cloning efficiency is 20%,less than some lung adenocarcinoma cell strain.