[Primary culture of the cells from Oncomelania hupensis liver].
OBJECTIVE
To develop a method for primary culture of cells from Oncomelania hupensis liver, and to observe the distribution of succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH) in the cultured cells.


METHODS
O. hupensis was anatomized to separate the liver. Livers were soaked in 0.2% benzalkonium bromide and washed by physiological saline containing antibiotics in turns. Cells from the liver were harvested by mechanical mulling and filtering. The isolated cells were then incubated with methods of the combination culture and standing suspension culture, respectively. The culture medium for the cells was a mixture of Medium 199 (50 ml), 0.3% lactoalbumin hydrolysate dissolved in a balanced salt solution (BBS, 30 ml), and fetal calf serum (FCS, 20 ml) containing a moderate amount of antibiotics (100 IU/ml penicillin, 100 microg/ml streptomycin and 50 microg/ml kanamycin) at pH 7.2-7.4 under the temperature of 26.5 degrees C. The cells were stained by using Giemsa and Pearson methods (for SDH and LDH respectively) to observe the shape of cultured cells and enzyme distribution in cells. The living and stained cells were microscopically observed.


RESULTS
Under microscope, the attached cells incubated with method of the combination culture showed round, elliptic, triangular and irregular shapes, with more round and elliptic cells. The size was approximately (4-16) microm x (6-20) microm in average. The clustered cells with an unclear nucleus and abundant and lucid cytoplasm were smaller than diffused cells with a large, obvious nuclei and less cytoplasm. Degeneration was observed after culturing for 5-7 days. The cultured cells could be divided into two types based on the color shown after Giemsa staining. The first type cells showed blue cytoplasm and mauve nuclei while the second type cells were opposite. There were blue granules in different sizes and shade in the cytoplasm after SDH and LDH staining. It was difficult for the cells to attach the wall of the culture flask using method of the standing suspension culture. The shape of the cultured cells were almost round with unclear nuclei, and the size was about (4-6) microm x (6-8) microm in average. The cells incubated with the standing suspension method were found to be contaminated after culturing for 3 days.


CONCLUSIONS
The combination culture method is suitable for primary culture of the cells from O. hupensis liver and the cells show activities of both SDH and LDH in cytoplasm.
Ye
Qing
Q
Department of Medical Parasitology and Research Laboratory of Schistosomiasis, Medical School of Wuhan University, Wuhan 430071, China.
Zhu
Jun-Yong
JY
Zhong
Qin-Ping
QP
Jiang
Ming-Sen
MS
Dong
Hui-Fen
HF
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation; D002478:Cells, Cultured; D003470:Culture Media; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000201:enzymology; D012908:Snails / Q000166:cytology* / Q000201:enzymology; D013385:Succinate Dehydrogenase / Q000378:metabolism
D003470:Culture Media; D007770:L-Lactate Dehydrogenase; D013385:Succinate Dehydrogenase