Introduction to PGHAM Cells
PGHAM cells are human pancreatic ductal epithelial cells that have been immortalized to retain characteristics of non-tumorigenic pancreatic tissue. Unlike many commonly used pancreatic cancer cell lines that are derived from metastatic or poorly differentiated tumors, PGHAM cells serve as a valuable model for studying the biology of normal or early-stage pancreatic epithelial cells. These cells are particularly useful for comparative studies with malignant lines to understand the molecular progression from normalcy to malignancy in pancreatic tissue. Their stable karyotype, epithelial morphology, and ability to form polarized monolayers make them ideal for modeling early oncogenic events, epithelial barrier function, and cellular responses to environmental stress.
Due to their non-cancerous origin, PGHAM cells do not typically exhibit the high proliferation rates or aggressive growth patterns seen in tumor-derived lines. This makes them especially suitable for mechanistic studies focusing on cellular homeostasis, innate immunity, DNA damage repair, and epigenetic regulation in pancreatic epithelia. They are also frequently used in toxicology research, testing the effects of xenobiotics, drugs, and gene modulators under near-physiological conditions. The retention of epithelial markers such as E-cadherin, cytokeratins, and tight junction proteins supports their use in epithelial integrity and permeability assays.
Importance of Transfection in PGHAM Cells
Transfection is an essential technique for functional genomics and molecular biology studies involving PGHAM cells. It allows researchers to introduce siRNA, miRNA, plasmid DNA, mRNA, or CRISPR-Cas9 constructs into the cells to probe gene function, analyze regulatory elements, or model disease states. Because PGHAM cells are non-tumorigenic and relatively stable, they provide a clean genetic background to assess the functional impact of single-gene perturbations without the confounding influences of extensive chromosomal abnormalities or high basal oncogenic signaling.
One of the primary challenges in transfecting PGHAM cells is their relatively low baseline proliferation and tight epithelial morphology, both of which reduce passive uptake of transfection complexes and limit the efficiency of standard lipid- or polymer-based reagents. Furthermore, these cells are sensitive to cytotoxicity and stress, which can activate non-specific stress pathways or cause irreversible damage to epithelial junctions. High-efficiency transfection in PGHAM cells, therefore, requires a specialized reagent that ensures efficient delivery while maintaining cellular health, polarity, and tight junction integrity.
Altogen PGHAM Transfection Reagent Overview
Altogen Biosystems has developed a proprietary transfection reagent tailored specifically for the unique requirements of PGHAM cells. The PGHAM Transfection Reagent is formulated with a blend of biodegradable cationic lipids and advanced polymer components that promote high-efficiency gene delivery while preserving cellular viability and phenotype. Unlike standard reagents that often result in off-target effects or epithelial disruption, Altogen’s reagent ensures minimal toxicity and is optimized for preserving junctional complexes and epithelial sheet architecture.
The reagent is compatible with DNA, siRNA, miRNA, and mRNA transfections, as well as CRISPR/Cas9-based genome editing tools. It forms stable, uniform nucleic acid complexes that rapidly internalize into PGHAM cells via endocytosis and achieve efficient endosomal escape. This results in high levels of gene expression or knockdown with minimal activation of cellular stress pathways. The reagent is designed for use with complete growth media, allowing for transfection under physiologically relevant conditions without media exchange or serum starvation.
Standard transfection efficiencies with plasmid DNA exceed 70%, while siRNA or miRNA-mediated knockdown consistently reaches 80–90%, with post-transfection viability typically above 90%. This high level of performance supports a broad range of experimental applications, including high-throughput screening, promoter activity assays, pathway mapping, and gene-environment interaction studies.
Transfection Protocol and Optimization
Optimal transfection in PGHAM cells with the Altogen reagent involves seeding cells at 50–60% confluence approximately 24 hours prior to transfection. Transfection complexes are prepared by separately diluting the nucleic acids and the transfection reagent in serum-free medium, followed by a 20-minute room temperature incubation to allow complex formation. These complexes are then added directly to the cells in complete medium.
Post-transfection, cells are typically monitored for gene expression, knockdown, or phenotypic changes within 24 to 72 hours. The reagent supports both transient and stable transfection protocols. For stable line generation, antibiotic selection (e.g., puromycin or G418) can begin 48 hours post-transfection, once cells have recovered and begun expressing the selection marker.
Variables that may require optimization include the DNA:reagent ratio, nucleic acid concentration, and incubation times. The reagent is sterile, endotoxin-free, and can be stored at 4°C for up to 12 months without loss of activity. Altogen provides detailed technical documentation to guide users in achieving optimal results in PGHAM cell transfection workflows.
Product Availability and Advantages
Altogen’s PGHAM Transfection Reagent is offered in multiple package sizes, including 0.5 mL, 1.5 mL, and 8.0 mL options, providing sufficient reagent for hundreds of transfections. It is manufactured under stringent quality control conditions to ensure batch-to-batch consistency and high performance.
Compared to general-purpose transfection reagents such as Lipofectamine or PEI, the Altogen PGHAM reagent demonstrates superior efficiency and cell viability. Its unique formulation reduces inflammatory responses, preserves epithelial integrity, and provides high reproducibility, making it ideal for applications in epithelial biology, early-stage cancer research, and toxicogenomics. For researchers focused on disease modeling, gene-environment interaction studies, or early-stage biomarker validation, this reagent offers unmatched performance and experimental reliability.
Research Applications
The Altogen PGHAM Transfection Reagent has been successfully used in a wide range of research applications. These include siRNA-mediated silencing of genes involved in epithelial cell polarity (e.g., ZO-1, claudin, occludin), overexpression of tumor suppressor genes (e.g., CDKN1A, TP53), and CRISPR-mediated knockout of regulatory genes involved in stress responses or DNA repair (e.g., ATM, ATR). The reagent is also used to transfect reporter constructs for monitoring transcriptional activity of key signaling pathways, including NF-κB, TGF-β, and Wnt.
PGHAM cells transfected with fluorescent or luminescent reporters exhibit high expression levels with minimal background noise, making them suitable for sensitive assay development and screening applications. Additionally, the reagent supports co-transfection of multiple plasmids or nucleic acids, allowing researchers to perform combinatorial gene perturbation studies or test complex regulatory networks.
For experiments involving epithelial permeability, cytokine secretion, or response to oxidative and ER stress, the Altogen reagent allows genetic manipulation without compromising cell health or junctional integrity. It is also compatible with 3D culture systems, organoids, and air-liquid interface cultures, further extending its versatility in translational and systems biology research.
Request the Altogen PGHAM Transfection Reagent
For high-efficiency, low-toxicity transfection of PGHAM pancreatic epithelial cells, request the Altogen PGHAM Transfection Reagent. Visit the product page to learn more and place your order.