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

Depositor: Professor Tony Burgess – Ludwig Institute for Cancer Research Ltd, Melbourne, Australia

References:

Whitehead RH, Jones JK, Gabriel A, Lukies RE. A new colon carcinoma cell line (LIM1863) that grows as organoids with spontaneous differentiation into crypt-like structures in vitro. Cancer Res 1987; 47:2683-9. PMID: 3567898.

Whitehead RH, Zhang HH, Hayward. Retention of tissue-specific phenotype in a panel of colon carcinoma cell lines: relationship to clinical correlates. Immunol Cell Biol. 1992; 70:227-36. PMID: 1427983.

Cellosaurus: CVCL_4434