Introduction

Colon cancer remains one of the leading causes of cancer-related deaths globally, necessitating advanced research methods to understand its pathology better and develop effective treatments. The human colon adenocarcinoma cell line, HT-29, serves as a vital model for investigating the efficacy of anti-cancer agents, particularly through proliferation assays. At Creative Biolabs, we adopt sophisticated HT-29-based proliferation assays to profile the antiproliferative potency and mechanisms of action of a variety of compounds, thereby contributing significantly to the oncology research landscape.

Human Colon Cell HT-29-based Proliferation Assay at Creative Biolabs

Creative Biolabs offers state-of-the-art HT-29-based proliferation assays to facilitate research in oncology. We provide several assay modes for global customers to suitable these projects, such as MTT assay, violet assay, and so on. By employing a highly standardized procedure, Creative Biolabs ensures the reproducibility and accuracy of our assays, providing high-quality data that meet stringent scientific standards.

Featured Assay Type

Luminescence-based assays possess high sensitivity and specificity, providing a dynamic range suitable for detecting subtle changes in cell proliferation. This assay is particularly advantageous for compounds with low potency levels, ensuring reliable detection and quantification of their effects on HT-29 cells. Leveraging the benefits of the luminescence-based assay, Creative Biolabs' HT-29-based luminescence proliferation assay services offer a powerful and reliable tool for oncology research and drug discovery.

Assay Details:

• Assay Type: Functional cell-based
• Detection Method: Luminescence
• Functional Mode: Antagonist
• Control Inhibitor: Staurosporine
• Measured Responses: Growth inhibition (GI50), total growth inhibition (TGI), and lethal concentration 50% (LC50)

Workflow

1. Cell Seeding: HT-29 cells are initially seeded in 96-well plates at a density of 1.5 × 10⁴ cells/ml, ensuring uniform cell distribution.
2. Incubation: The cell culture plates are incubated overnight in a controlled environment (37°C, 5% CO₂) to allow cell attachment and acclimatization.
3. Compound Addition: Post incubation, test compounds or vehicle controls are administered to the cells, followed by a further 72-hour incubation.
4. Detection: After the incubation period, cell proliferation is assessed using a luminescence microplate reader. Growth inhibition (GI50), total growth inhibition (TGI), and lethal concentration (LC50) are calculated based on luminescent signals.
5. Analysis: Data is meticulously analyzed to generate dose-response curves and infer compound efficacy.

Benefits of Our HT-29-based Proliferation Assay

• Precision: Utilization of high-throughput, image-based assay techniques for accurate cell number and phase distribution determination.
• Scalability: Ability to adapt protocols to high-density 96 or 384-well plate formats for extensive compound screening.
• Customization: Tailored assay parameters to fit specific client needs, encompassing various dosages and treatment durations.
• Reliability: Employment of robust assay conditions, including the use of control inhibitors, to ensure consistency and reproducibility of results.

Scientific Backing

The application of HT-29 cells in proliferation assays is supported by extensive scientific research. For example, the use of HT-29 cells-based proliferation assay by the MTT method enables to study the metastasis signaling pathway.

Fig.1 Increased proliferation of metastatic colon carcinoma cells after DEX treatment analyzed by MTT assay.¹

Frequently Asked Questions

Q1: How does the luminescence detection method work?

A1: The luminescence detection method measures cellular ATP levels, which are directly proportional to cell viability. The light emitted during the ATP-dependent reaction with luciferase provides a quantifiable signal correlating with cell number.

Q2: Why is the HT-29 cell line chosen for proliferation assays?

A2: HT-29 cells offer a reliable model system due to their similarity to human colonic epithelial cells, robust proliferation characteristics, and predictable response to chemotherapeutic agents, making them ideal for studying colon cancer cell proliferation and drug efficacy.

By integrating comprehensive HT-29-based proliferation assays, Creative Biolabs exemplifies a commitment to advancing colon cancer research through scientific excellence and innovative methodologies.

Reference

1. Tian, Dan, et al. "Increased glucocorticoid receptor activity and proliferation in metastatic colon cancer." Scientific Reports 9.1 (2019): 11257. Distributed under Open Access License CC BY 4.0, without modification.

For Research Use Only | Not For Clinical Use