A549 Cell Line Transfection Protocol

The A549 cell line was established in 1972 by D.J. Giard et al, through an explant culture of adenocarcinomic lung tissue of a 58-year-old Caucasian male. A549 cells are hypotriploid alveolar basal epithelial cells. This adenocarcinomic cell line is categorized as a non-small-cell lung carcinoma (NSCLC), which tends to be less aggressive and spread less quickly than small cell lung carcinoma (SCLC) but proves to be more common, accounting for 85-88% ofall cases of lung cancer. The A549 cell line is widely used as a model of lung adenocarcinoma, as well as an in vitro model for type II pulmonary epithelial cells.

Cancer research is an area of scientific inquiry that seeks to identify the causes of cancer, find new treatments and develop strategies to prevent the disease. One type of cancer that is particularly difficult to treat is lung cancer. Lung oncology is one of the leading causes of cancer-associated deaths worldwide, and researchers are constantly working to find new ways to diagnose, treat and prevent the disease. Lung cancer is the leading cause of cancer death in both men and women, and the five-year survival rate for the disease is below 20 percent. The A549 cell line is a widely used human lung adenocarcinoma cell line that was derived from a primary lung tumor. A549 cells are highly proliferative and have been used in a wide variety of research applications.

Cytogenetic Information

The A549 cell line is hypotriploid with a modal chromosome number of 66, which occurs in 24% of cells. Modal numbers of 64 and 67 is relatively common with higher ploidies occurring at an infrequent rate (0.4%). A549 cells are positive for keratin by immunoperoxidase staining. The cells are able to synthesize lecithin and contain a high
percentage of unsaturated fatty acids, which are utilized by the cytidine-diphosphocholine pathway and important for the maintenance of membrane phospholipids in cells.

Morphology and Characteristics of A549 Cells

A549 cells have been well characterized over the years and are valuable to researchers that routinely use them as in vitro and in vivo models. These cells are squamous in nature and are responsible for the diffusion of substances such as water and electrolytes across the alveoli of lungs. A549 cells are human alveolar basal epithelial cells that grow adherently as a monolayer in vitro and serve as suitable transfection hosts.

A549 cells are epithelial cells, meaning they are cells that line the surfaces of organs and tissues. A549 adenocarcinoma cells are grown in a laboratory in a nutrient-rich DMEM culture medium. When they are grown in this way, they form a monolayer (single layer of cells). A549 cells became a golden standard for lung cancer research model and now commonly used in cell culture studies as a model for studying human lung cancer.

A549 Cell Line Derived Xenograft Mouse Model

The A549 non-small-cell lung carcinoma (NSCLC) cell line is cultured and then inoculated in mice to create the A549 CDX mouse model (Cell Line Derived Xenograft). The A549 CDX model is one of the most used xenograft lung cancer models by researchers for studying SoC (standard of care) and novel therapeutics (i.e. gefitinib, erlotinib, paclitaxel. lapatinib) due to overexpression of HER-2 and EGFR receptors. Typical end of study results are comprised of tumor volume measurements, body weight tracking or animal survival.

The following options are typically available for an A549 CDX xenograft model:
• Dosing frequency and duration of dose administration
• A549 Tumor Growth Delay
• A549 Tumor Growth Inhibition
• Dosing routes:
    ·Intravenous
    ·Intratracheal
    ·Continuous infusion
    ·Intraperitoneal
    ·Intratumoral
    ·Oral gavage
    ·Topical
    ·Intramuscular
    ·Subcutaneous
    ·Intranasal
    ·Cutting-edge micro-injection techniques and pump-controlled IV injection
• A549 tumor immunohistochemistry
• Cell engraftment sites:
    ·Orthotopic transplantation
    ·Tail vein injection
    ·Left ventricular injection for metastasis studies
    ·Injection into mammary fat pad
    ·Intraperitoneal injection
• Blood chemistry analysis
• Toxicity and survival (optional: performing a broad health observation program)
• Gross necropsies and histopathology
• Imaging studies: Fluorescence-based whole body imaging, MRI
• Positive control group employing cyclophosphamide, at a dosage of 50 mg/kg administered by intramuscular injection to the control group daily for the study duration
• Lipid distribution and metabolic assays

Other lung cancer xenograft models available here

Publications

• Chemokine production by A549 cells in response to Mycobacterium tuberculosis: In this study, A549 cells were infected with several strains of M. tuberculosis and then their chemokine production was measured. In addition, the intracellular growth of the A549 cells was measured and compared with the chemokine production. LINK
• Characterization of an in vitro model of Type II alveolar epithelium using A549 cells: This study recorded the absorption and transport of different markers by A549 cells. Cationized ferritin (nonspecific absorptive marker) and transferrin (receptor-mediated
endocytic marker) were absorbed by the A549 cells, whereas Lucifer yellow (fluid-phase marker) was not. Fluorescein-transferrin caused the A549 monolayers to polarize, thus allowing a greater amount of transferrin to be transported out of the cells, and these results indicated the possibility of using A549 cells to study the
metabolic and macromolecule contributions of alveolar Type II cells to mechanisms of drug delivery at the pulmonary epithelium. LINK
• Apoptosis of A549 triggered by evodiamine through MTDH-dependent signaling pathway: This study investigated the anti-tumor effect of evodiamine on the A549 cell line and the inhibitory effect of evodiamine on MTDH (Metadherin), an oncoprotein that can promote the survival and spread of lung cancer cells when
overexpressed. Staining assays and a Western blotting analysis showed that evodiamine suppressed the proliferation of A549 cells by inhibiting MTDH expression and activating apoptosis, which indicates that evodiamine exhibits the potential to be
developed as a cancer therapeutic. LINK
• Observation of differentiation in A549 cells: Monolayer cultures of A549 cells were maintained for up to three weeks, and examined using an immunoperoxidase technique. This technique used antibodies to detect surface-associated glycoproteins,
which bound differently to certain A549 cells, indicating localization of the glycoproteins. LINK

Links

A-549 human lung carcinoma (DSMZ)
siRNA Delivery – In Vivo Transfection Kits
CRO Pre-clinical Research Services: Xenograft animal models
GLP-compliant Cell Banking services
Generation of Stably Expressing Cell Lines in 28 Days
Stable RNAi Cell Line Generation: Stable Gene Knockdown
In Vivo siRNA Delivery: Tissue-targeted siRNA
Encapsulation of Protein, RNA, mRNA, and DNA Molecules into Liposomes
Purchase A549 cell line (ATCC)
A549 Cell (Wikipedia)

Proven transduction protocols for the most popular cancer cell lines and varieties are introduced. These arrangements assist you in receiving the outcome you hope to get the first time with few expectations. Search for your specific cell line protocol for breast, lung, prostate, or liver cancer. These new protocols help you with an easy plan to help satisfy the high transduction effectiveness and low toxicity for your special cell line when paired with Lipofectamine 3000 transduction regimen. The protocols go over growing cells, culture conditions, and passing through a successful transduction outcome.

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