BACKGROUND:There is a certain cel subset in esophageal cancer tissues, with certain invasive and metastatic properties, which is closely related to the clinical therapeutic effect on tumors.
OBJECTIVE:To isolate tumor stem cel spheres in human esophageal carcinoma cel lines KYSE-150 and TE-1 and to analyze their proliferation and invasion ability. METHODS:KYSE-150 and TE-1 cels were cultured in serum-free medium to observe the formation of cel spheres. Cel proliferation and invasion were detected using MTT and Transwel chamber culture. Surface markers of cels were detected using flow cytometry. RESULTS AND CONCLUSION: Cel spheres that were stably subcultured were obtained from KYSE-150 and TE-1 cels cultured in serum-free medium. The proliferation and invasion abilities of cel spheres were significantly stronger than those of parent cels (P < 0.05). The number of CD44+, CD271+ and CD44+CD271+ cels in TE-1 and KYSE-150 cel spheres was significantly higher than that in the TE-1 and KYSE-150 parent cels (P < 0.05). These experimental results show that cel spheres isolated from human esophageal carcinoma cel lines TE-1 and KYSE-150 have tumor stem cel properties as wel as strong proliferation and invasion abilities. And moreover, CD44 and CD271 can be used as important surface markers of esophageal carcinoma stem cels. Cite this article:Wang YL, Wang ZM, Wang Y, Tao YP, Han GY.Culture, differentiation, proliferation and invasion of tumor stem cels in human esophageal carcinoma cel lines KYSE-150 and TE-1. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):55-59.

OBJECTIVE: To analyze and characterize the separation effectiveness of self-constructed asymmetrical flow field-flow fractionation system on proteins and lipoproteins, to achieve the optimization of the experimental conditions when separating lipoproteins by orthogonal design test and to investigate the carrier viscosity's influence on separation effectiveness. METHODS: The evaluation of asymmetrical flow field-flow fractionation separation capacity was conducted by using two standard proteins (carbonic anhydrase and thyroglobulin). Under the optimized separation conditions of carbonic anhydrase and thyroglobulin, the channel actual thickness (after assembling, the actual thickness of separation channel was less than initial thickness) was calculated by the analytes' elution time based on the hydrokinetic theory. With orthogonal design test the optimized experimental conditions were studied and statistical analysis was carried on to find out the factors with statistical significance which needed further exploration. RESULTS: According to the hydrodynamics principle and Stoke's function, the channel actual thickness was measured to be 164 μm by separating the two standard proteins, carbonic anhydrase and thyroglobulin, under proper experimental conditions. By the optimization based on orthogonal design test, base-line separation (the resolution had to be higher than 1.50) was achieved. The resolutions of the two experiments were 1.61 and 1.58. According to previous study/ pre-study and supporting theory, in the orthogonal design test, the total 5 factors were integrated for comprehensive investigation: the total flow rate (3.00, 3.50, 4.00, 4.50 mL/min), focus time (3.00, 3.50, 4.00, 4.50 min), transition time (0.5, 1.0, 1.5, 2.0 min), pH of the carrier fluid(6.8, 7.00, 7.20, 7.40) and viscosity of the carrier fluid hydroxypropylmethylcellulose concentration: 0.00%, 0.03%, 0.06%, 1.00%). Among the 5 factors, viscosity was found to have the statistical significance on separation effectiveness which was further investigated. The resolution of high density lipoprotein and low density lipoprotein was increased by the increasing viscosity which also caused more obvious negative spikes. CONCLUSION The separating capacities of self-constructed asymmetrical flow field-flow fractionation system on lipoproteins were verified to be effective and an optimized experimental condition was found to achieve the base-line separation of high density lipoprotein and low density lipoprotein. Viscosity of the carrier fluid was proved to have the statistical significance on lipoprotein separation.
Fractionation, Field Flow ; Lipoproteins ; Lipoproteins, LDL