久久精品99国产I国产精品国产亚洲精品看不卡I日韩区视频I国产丝袜在线I久久精品小视频Iadn—256中文在线观看I夜夜高潮夜夜爽国产伦精品I亚洲女裸体

This cell line was developed from a wedge biopsy of a liver metastasis of a pancreatic ductal carcinoma.

The cells can be lysed completely in vitro by autologous and allogeneic lymphokine activated killer (LAK) cells in the presence of interleukin 2 (IL-2). They are relatively insensitive to autologous and allogeneic natural killer (NK) cell lysis.

The cells produce CEA, and grow in orthotopic and heterotopic positions in nude mice within 21 days at 100% frequency (5/5).

Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. 

1. Remove and discard culture medium. 
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum, which contains trypsin inhibitor. 
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and place at 37°C to facilitate dispersal. Observe cells within 5 to 10 minutes under an inverted microscope 
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. 
5. To remove trypsin-EDTA solution, transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. 
6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Recommended seeding densities of 5 x 10³ to 4 x 10⁴ viable cells/cm² 
7. Place culture vessels in incubators at 37°C. Note: subcultures can also be prepared by scraping the cells from the floor of the flask. Add fresh medium, aspirate the scraped cells to disperse them and dispense into new flasks.

Drucker BJ, et al. A new human pancreatic carcinoma cell line produced for adoptive immunotherapy studies with lymphokine-activated killer cells in nude mice. In Vitro Cell. Dev. Biol. 24: 1179-1187, 1988. PubMed: 3264833