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DLD-1 Cells: Applications and Insights of DLD-1 Cells in Colorectal Cancer Research

DLD-1 is a human colorectal carcinoma cell line commonly employed in cancer research. It is a valuable tool for investigating various aspects of colon cancer, including tumour development, progression, underlying molecular mechanisms, and response to therapeutic agents. Thus, DLD-1 is outstanding in understanding disease and developing potential treatment strategies.

This article will provide important information about the DLD-1 cell line you should know before working with it. It will cover:

  1. General characteristics and origin of DLD-1 cells
  2. Culturing information of DLD-1 cell line
  3. DLD-1 cell line: Advantages & Limitations
  4. Research applications of DLD-1 cell line
  5. Research publications on DLD-1 cells
  6. Resources for DLD-1 cell line: Protocols, Videos, and More

1.      General characteristics and origin of DLD-1 cells

Before working on a cell line, the first consideration of a researcher is its origin and general attributes. This article section is a detailed introduction to the DLD-1 cell line. It will cover the following frequently asked questions: What are DLD-1 cells? What is a DLD-1 cancer cell line? What are the DLD-1 cell line mutations? What is the size of a DLD-1 cell? What is the morphology of DLD-1 cells?

  • DLD-1, a colon cancer cell line, originated from the large intestine of a 67-year-old male patient with colorectal adenocarcinoma. It was isolated by D.L. Dexter and colleagues during 1977-1979. This cell line shares the same origin with HCT-8, HCT-15, and HRT-18 cell lines.
  • Like colorectal cancers, DLD-1 cells possess a microsatellite instability phenotype [1].
  • DLD-1 mutations include KRAS, P53, BRAF, BRCA1 and BRCA2. Due to DLD-1 P53 status, DLD-1 BRCA, DLD-1 BRCA2, and DLD-1 KRAS mutation, these cells are considered ideal for cancer studies.
  • These cells possess an epithelial-like morphology.
  • The size of the DLD-1 cell line is approximately 15 µm.
  • The human colorectal cancer cell line DLD-1 possesses a pseudodiploid karyotype. Around 86% of cells possess a modal chromosome number of 46. However, polyploidy may also exist in 17.1% of the cell population.

3D rendering of a CT colonography or CT scan of the large intestine displayed on the screen for the diagnosis of large bowel cancer, colon cancer, and colorectal cancer.

2.      Culturing information of DLD-1 cell line

Before culturing any cell line, you must know its basic cell culture requirements. This section will highlight the important points you must learn before commencing work on the DLD-1 cell line. This includes: What is DLD-1 doubling time? What is the DLD-1 culture medium? What is the seeding density of DLD-1 cells? How do you culture DLD-1 cells?

Key Points for Culturing DLD-1 Cells

Doubling Time:

The DLD-1 doubling time is approximately 15 hours.

Adherent or in Suspension:

The DLD-1 is an adherent human colon cancer cell line. Cells grow into monolayers.

Seeding Density:

DLD-1 cells are seeded at a density of 1 to 2 x 104 cells/cm2. Adherent cells are washed with 1x phosphate buffer saline (PBS) and incubated with passaging solution (Accutase) at an ambient temperature. After incubation of 8 to 10 minutes, detached cells are added to the culture medium and centrifuged. The cell pellet is then carefully resuspended, and cells are poured into new culture vessels containing growth medium.

Growth Medium:

RPMI 1640 with medium supplements, including 10% fetal bovine serum FBS, 2.1 mM stable Glutamine, and 2.0 g/L NaHCO3 is used to culture DLD-1 cells. Media should be replaced 2-3 times per week. 

Growth Conditions:

DLD-1 cultures are maintained in a 37°C humidified incubator with a 5% CO2 supply.


DLD-1 cells are stored in the vapour phase of liquid nitrogen or an electric ultra-low temperature freezer (at below -150°C temperature).

Freezing Process and Medium:

CM-1 or CM-ACF freezing media can be used to freeze the DLD-1 cells. A slow freezing process is always recommended, allowing a gradual 1°C decrease in temperature, thus preventing cells from shock and protecting their viability.

Thawing Process:

Frozen cells are thawed in a water bath pre-set at 37 °C temperature for 40 to 60 seconds until a small ice clump is left. Afterwards, cells are added to a fresh medium and centrifuged. This step is necessary to remove freezing media components from cells. The collected cells are resuspended in a growth medium and dispensed into the flask for growth.

Biosafety Level:

The DLD-1 cell line is maintained in the biosafety level 1 laboratories.

DLD-1 cells observed at 10x and 20x magnification, revealing their characteristic epithelial morphology.

3.      DLD-1 cell line: Advantages & Limitations

In this section of the article, we will review the advantages and limitations of the DLD-1 colon cancer cell line.


The main advantages of the DLD-1 cell line are:

Colorectal Cancer Model

DLD-1 cells possess mutations commonly found in colorectal cancer patients, such as DLD-1 KRAS mutation, DLD-1 BRCA, and DLD-1 BRCA2. This relevance makes them a clinically appropriate cell model for studying this disease.

Tumorigenic Cell Line

DLD-1 cells are tumorigenic. They can cause tumours when injected into nude mice. Therefore, researchers employ these cells to develop xenograft tumour models that help investigate tumour formation, development, and progression in vivo.



The limitations associated with the DLD-1 cells are:


DLD-1 colon cancer cell populations exhibit genetic and phenotypic heterogeneity. This may influence the generalizability of research outcomes.


4.      Research applications of DLD-1 cell line

DLD-1 cell line offers a range of applications in cancer research. Some prominent applications are discussed in this section.

  • Cancer study: DLD-1 cells serve as a valuable in vitro model for understanding the molecular mechanisms underlying colorectal cancer growth, development, and progression. Moreover, they are used for studying gene expression, signalling pathways, and the influence of specific genetic mutations on cancer cell behaviour. Research conducted in 2022 observed that PI3K/Akt/YAP signaling cascade regulates the migration and invasion of the DLD‑1 colorectal cancer cell line [2].
  • Drug development: Researchers employ the DLD-1 cell line to test the effectiveness of anti-cancer drugs and evaluate their potential for colon cancer treatment. This aids in drug development and combating disease efficiently. A study conducted in 2022 used the DLD-1 cancer cell line and explored the anti-cancer activity of silicon phthalocyanines, chemically synthesized compounds. The study outcomes suggested these compounds as potent anti-cancer agents [3]. Interestingly, another research found the anti-cancer potential of a probiotic-derived P8 protein using DLD-1 cells [4].

5.      Research publications on DLD-1 cells

This section will highlight a few interesting and frequently cited publications featuring the DLD-1 colon carcinoma cell line.

Flavonoid glycoside diosmin induces apoptosis and cell cycle arrest in DLD-1 human colon cancer cell line

This publication is in the Journal of Biologically Active Products from Nature (2022). The study found that diosmin, a flavonoid, induced cell death in DLD-1 cells and also caused cell cycle arrest; thus, it can be a potential anti-cancer agent.

Bromelain effectively suppresses Kras-mutant colorectal cancer by stimulating ferroptosis

This study in Animal Cells and Systems (2018) proposed that bromelain, an enzyme, represses DLD-1 KRAS mutation and exerts potent cytotoxic effects.

Bergapten induces G1 arrest and pro‐apoptotic cascade in colorectal cancer cells associating with p53/p21/PTEN axis

This research article was published in 2019 in the “Environmental Toxicology” journal. The study explored the anti-tumor effects of a natural compound, Bergapten, using a DLD-1 corectal cell line.

Eco-Friendly Fabrication of Nanocurcumin/Nano Iron Oxide Composite: Enhanced In Vitro Anticancer Activity Against DLD-1 Cell Lines

This study in the Journal of Inorganic and Organometallic Polymers and Materials (2023) found that curcumin/iron oxide nanocomposites are highly effective against DLD-1 colorectal cancer cells.

Epibrassinolide activates AKT to trigger autophagy with polyamine metabolism in SW480 and DLD-1 colon cancer cell lines

This research article was published in 2020 in the Turkish Journal of Biology. The study proposed that a natural product, epibrassinolide-mediated activation of AKT in DLD-1 cells, stimulates autophagy related to polyamines.

6.      Resources for DLD-1 cell line: Protocols, Videos, and More

The following are a few resources on DLD-1 cells:

  • DLD-1 transfection: This video tutorial is a comprehensive guide about DLD-1 in vitro transfection protocol.
  • DLD-1 cells: This research paper briefly describes DLD-1 cell culture requirements, including media and culture conditions. Besides, it may help you learn the transfection protocol used for DLD-1 cells.

The following links contain the protocol for culturing DLD-1, human-derived colon cancer cells.

  • DLD-1 cell line: This website will provide basic information about the DLD-1 cell line. It will help you learn different DLD-1 cell culture protocols, including subculturing and handling of cryopreserved and proliferating cultures.


  1. Gavrilas, L.I., et al., Pro-apoptotic genes as new targets for single and combinatorial treatments with resveratrol and curcumin in colorectal cancer. Food & function, 2019. 10(6): p. 3717-3726.
  2. Takeda, T., et al., PI3K/Akt/YAP signaling promotes migration and invasion of DLD1 colorectal cancer cells. Oncology Letters, 2022. 23(4): p. 1-9.
  3. Farajzadeh, N., et al., Anticancer activity of novel silicon phthalocyanines against the colorectal adenocarcinoma cell line (DLD-1). New Journal of Chemistry, 2022. 46(41): p. 19863-19873.
  4. An, B.C., et al., Anti-Cancer Roles of Probiotic-Derived P8 Protein in Colorectal Cancer Cell Line DLD-1. International Journal of Molecular Sciences, 2023. 24(12): p. 9857.


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