Liver cancer cell system from liver cancer portal vein tumor thrombus of human and system establishing method thereof
A technology of liver cancer cell line and portal vein, applied in the field of liver cancer cell line and its establishment
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Embodiment 1
[0026] Establishment of HCC cell line derived from portal vein tumor thrombus of human HCC
[0027] 1. Materials
[0028] 1. Two different cell culture media and their preparation methods
[0029] 1.1 Culture solution D / F: DMEM / F12 (1:1) (GIBCO, USA), 10% (v / v) fetal bovine serum (Biochrom AG, USA), EGF (Epidermal Growth Factor, epidermal growth factor) 20ng / ml (U.S. Sigma Company), 5ug / ml insulin, 0.30g / L L-glutamine, 100U / L penicillin, 100ug / L streptomycin;
[0030] 1.2 Culture solution DMEM: high-sugar DMEM medium (GIBCO, USA), 10% (v / v) fetal bovine serum (Biochrom AG, USA), 0.30g / L L-glutamine, 100U / L penicillin, 100ug / L Streptomycin.
[0031] 2. Other cell culture materials
[0032] Cell sieve (BD Company, U.S.); program freezing box (NALGENE Company, U.S.); 0.05% type I collagenase solution (Sigma Chemical Company, St.Louis, MO); trypsin digestion solution: 0.25% trypsin (GIBCO, U.S. company)+0.02% EDTA, pH value is 7.3; cell cryopreservation solution (DMEM:FBS:D...
Embodiment 2
[0054] Biological characteristics of liver cancer cell lines derived from portal vein tumor thrombus of human liver cancer
[0055] In this example, conventional methods were used to detect the biological characteristics of the cell line CSQT-1 with the deposit number CCTCC NO: C200814. Methods as below:
[0056] 1. Experimental method
[0057] 1. Morphology
[0058] (1) General shape
[0059] Two days after cell inoculation, the growth morphology of the population cells was observed.
[0060] (2) light mirror
[0061] The morphological structure and growth characteristics of the cultured living cells were observed under an inverted phase-contrast microscope.
[0062] (3) electron microscope
[0063] The 40th passage cells were taken, and the ultrastructure of the cells was observed by scanning and transmission electron microscopy.
[0065] (1) Drawing of cell growth curve and cell population doubling time
[0066] Take well-grown logarithmic...
Embodiment 3
[0111] Culture medium D / F (1)
[0112] DMEM / F12 (1:1) (GIBCO, USA)
[0113] 10% (v / v) fetal bovine serum (Biochrom AG, USA)
[0114] EGF (Sigma, USA) 20ng / ml
[0115] Insulin 5ug / ml
[0116] L-Glutamine 0.30g / L
[0117] Penicillin 100U / L
[0118] Streptomycin 100ug / L
[0119] Preparation method: The culture medium D / F can be prepared according to the above concentration.
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