Colo205
The Colo205 is a laboratory instrument designed for the analysis and measurement of various samples. It utilizes advanced techniques to provide accurate and reliable data. The core function of the Colo205 is to perform analytical tasks, but its specific intended use is not included in this factual description.
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63 protocols using «colo205»
Propagation of Colorectal Cancer Cell Lines
Propagation of Colorectal Cancer Cell Lines
Culturing Colo-205 and KHYG-1 Cells
KHYG-1 cells were cultured under identical conditions as Colo-205, supplemented with an additional 100 U/mL of recombinant human interleukin-2 (Peprotech, 200-02). Both cell types were seeded in a 90 × 20 mm cell culture dish (SPL, 20101). The medium was changed every 3-4 days.
Cell Line Culture and Authentication
from the American Type Culture Collection via LGC Standards. Human
normal lung fibroblasts (MRC-5), alveolar basal epithelial cell adenocarcinoma
(A549), colorectal adenocarcinoma (Colo205), hepatocellular carcinoma
(HepG2), breast adenocarcinoma (MCF7), and colorectal adenocarcinoma
(SW620) and its MDR variants55 (link) were cultured
in standard conditions (37 °C, 5% CO2, 100% relative
humidity) in high glucose DMEM medium supplemented with GlutaMax,
HEPES (ThermoFisher Scientific) and 10% fetal bovine serum (EURx,
Poland). All cell lines were tested for Mycoplasma contamination using a MycoProbe mycoplasma detection kit (R&D
System).
CRC Cell Line Genetic Manipulation
To knock down HIF1A-AS2 in CRC cells, short hairpin RNAs (shRNAs) were cloned into the pGLV3/H1/GFP/puro vector (sh-HIF1A-AS2#1/sh1 and sh-HIF1A-AS2#2/sh2) (Shanghai Fanxu Technology). Full-length HIF1A-AS2 was cloned into pCDH-CMV-MCS-EF1-copGFP-T2A-Puro plasmid (ex-HIF1A-AS2/HIF1A-AS2) to overexpress HIF1A-AS2. HEK-293T cells were transfected with the knockdown/overexpressed plasmids, psPAX2, and pMD2.G, using a transfection reagent. After 48 h, the culture medium of HEK-293T cells (containing lentivirus particles encapsulating knockdown/overexpression constructs) was harvested, filtered using a percolator whose bore diameter is 0.45 μm (Millipore, USA), and applied to transfect CRC cells to knock down or overexpress HIF1A-AS2. Stable cell lines were selected using a culture medium containing 5 µg/mL puromycin. Short hairpin RNAs against FOXC1 (sh-FOXC1) and relevant overexpression plasmids (ex-FOXC1), were also designed. SP1, FOXP3, E2F1, wild-type and mutant HIF1A-AS2 were cloned into pcDNA3.1 vector. siRNAs against FOXP3, SP1, E2F1, and the mimics or inhibitors of miR-141-3p were transfected into CRC cell line to change the expression of relevant genes. Supplementary Table
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