LIM 1863 cell line细胞株-BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

Description: LIM1863 is a cell line derived from a sporadic colorectal cancer (CRC), from a 74-year-old female patient. The cells grow as organoids and are organized around a central lumen. Both morphologically mature columnar and goblet cells are present in the organoids.



Description Key Words: Sporadic Colorectal Cancer (CRC), carcinoma of ileoceacal valve



Also Known As: No other names



Organism: Human (Homo sapiens)



Strain: N/A



Tissue: Colon



Growth Properties: Suspension



Morphology: Cells grow as floating organoids.



Growth Medium: RPMI 1640 (with 2mM L-Glutamine + 25mM HEPES) + 10% Foetal Calf Serum (FCS) + Insulin 0.6μg/ml, Hydrocortisone 1μg/ml, 1-Thioglycerol 10μM.



Resuscitation: Remove protective cryoflex layer around the ampoule prior to thawing. Thaw the ampoule by gently agitating in a 37°C waterbath; thawing should be rapid (around 2 minutes). A centrifugation step to remove the cryoprotectant after thawing is necessary for this cell line.

Seed the thawed cells into a T75 flask, the cells grow as floating organoids and are frozen in clusters. Do not count at thaw. Split 4-6 days after resuscitation.



Subculturing Procedure:



Medium Renewal: 3 times each week.



Subcultivation Ratio: 1:4, Split once a week at this optimal ratio.

To subculture, disperse the organoids into smaller clusters by vigorous pipetting and then split 1:4 to seed.

Note: This cell line does not tolerate low density. If there are too few organoids in the flask to grow well, they may need to be concentrated into a smaller volume.

To obtain an estimated cell count, remove an aliquote of the cells suspension and after PBS wash, use 0.05% Trypsin/EDTA at 37°C for 5 minutes with mechanical dispersing before cell count. (Seeding density 2-4 x104 cells/cm2).



Culture Conditions: Cells maintained at 37°C and 5% CO2.



Cryoprotectant Medium: 10% DMSO + 90% FCS



Handling Procedure for Frozen Cells: Upon receipt, frozen ampoules should be transferred directly to liquid nitrogen storage without delay, if not to be used immediately. Storage at -80°C may result in loss of viability.



Additional Information:

LIM1863 will adhere in response to TGF-beta
Truncated APC (stop at aa1506)
Mutated beta-catenin
A33 positive

Medium:
Growth Condition:

BioVector NTCC Inc.
www.biovector.net