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hTERT-HPNE-菌种及细胞-北纳生物

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基本信息
资源编号 BNCC239243
资源名称 hTERT-HPNE
种属 Homo sapiens, human│Pancreas, duct
类型 intermediary cells formed during acinar-to-ductal metaplasia
形态 Epithelial-like
提供形式 frozen
安全等级 2
培养方法
培养基 The base medium for this cell line is: 75% DMEM without glucose (Sigma Cat#. D-5030 with additional 2 mM L-glutamine and 1.5 g/L sodium bicarbonate) 25% Medium M3 Base (Incell Corp. Cat# M300F- 500) To make the complete growth medium, add the following components to the base medium: fetal bovine serum 5% (final conc.) 10 ng/ml human recombinant EGF 5.5 mM D-glucose (1g/L) 750 ng/ml puromycin
传代方法 Volumes used in this protocol are for 75 cm2. flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37&deg;C to facilitate dispersal. Add 2.0 to 3.0 mL of complete growth medium and aspirate cells by gently pipetting. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 4 x 103 to 6 x 10 3 viable cells/mL is recommended. Incubate cultures at 37&deg;C. Subculture when reaching a concentration between 5 x 104 and 6 x 104 cells/cm2. <br>Subcultivation Ratio: 1:8 - 1:12 twice weekly<br>Medium Renewal: Every 2 to 3 days Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
生长条件 Temperature: 37&deg;C<br>Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
生长特性 adherent
存储条件 Liquid nitrogen vapor phase;Freeze Medium: 95% FBS; 5% DMSO
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