An inquiry to experimental conditions and cell-counting assays in reconstituted basement membrane invasion experiment in vitro
- VernacularTitle:体外重组基底膜侵袭实验条件及计数方法的探讨
- Author:
Nan LI
;
Peng SHANG
;
Airong QIAN
- Publication Type:Journal Article
- Keywords:
reconstituted basement membrane;
invasion;
chemoattractant;
staining
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the most suitable chemostatic condition and a method of cell-counting in reconstituted basement membrane invasion experiment in vitro. Method Reconstituted basement membrane invasion experiment was employed, and conditioned medium, fibronectin (FN), and a combination of conditioned medium and FN were respectively used as chemoattractants was placed in the lower compartment of the chamber. After the filters were stained with hematoxylin and eosin (HE), two kinds of cell-counting methods were employed. One was random cell-counting to count the cells in 5 fields infiltrated in the different areas of the filter membrane under 40?magnification, and the other was whole cell-counting of the infiltrating cells on the whole filter membrane with 4? magnification and image analysis system. Results The number of infiltrating cells in the presence of the chemoattractants was much larger than that of the control group. The number of infiltrating cells of the conditioned medium group was larger than that of FN group. The numbers of infiltrating cells in the combination of conditioned medium and FN groups were significantly larger than those of conditioned medium and FN alone groups. The accuracy of method of random cell-counting under 40? magnification was poorer than that of whole cell-counting with image analysis system. Conclusion Chemoattractants exert strong effect on the tumor cells invasion, and different chemoattractants show different chemotactic abilities. By using image analysis system to count all the infiltrative cells of the whole filter membrane is an objective and accurate cell-counting method.
