Explore our iPSC assay solutions: trilineage differentiation, lentiviral protocols, morphology & key pluripotency markers, and much more. Learn about how our iPSC assay services can accelerate your research.
iPSC Assays
Why iPSC Assays are Essential for Research
Induced Pluripotent Stem Cells (iPSCs), with their ability to differentiate into all cell types of the body, have quickly become a cornerstone of modern biotechnological research. Compared to Embryonic Stem Cells (ESCs), which are also pluripotent, iPSCs present a more ethical avenue, allow for sourcing multiple cell types from a single patient, and offer the unique advantage of deriving cells from disease-affected individuals, further enriching their potential in research and therapy.
Expanding the Horizons: iPSCs in Cell Engineering
Beyond their foundational role in research, iPSCs are increasingly being utilized for cell engineering techniques to manipulate genes and integrate reporters. Common approaches involve lentiviral transduction, CRISPR/Cas9, or electroporation. Elixirgen Scientific recommends the use of lentiviral transduction for quick, stable, and robust integration, with the added benefit of drug selection capability to purify engineered populations. The primary objectives of these engineering efforts are usually fluorescent reporter gene integrations or disease-specific gene knock-downs. These modifications are designed to simplify and improve the robustness of downstream assays after differentiation. Please refer to our application note for information on using lentiviral transduction with iPSC-derived excitatory neurons.
Ensuring Quality and Consistency
With multiple methods of generating and manipulating iPSC, results may vary; thus it is imperative to characterize and ensure the quality of the iPSC lines before conducting research with them. Elixirgen Scientific employs a combination of advanced cellular assays and detailed molecular analyses as a part of our own internal production process, quality control processes, and as a service to our customers.
Overview of iPSC Assays
Assays play a pivotal role in iPSC characterization. They are sophisticated techniques used to analyze and validate the quality, pluripotency, identity, and differentiation potential of iPSCs. Assays can range from simply observing iPSC morphology to more complex tests designed to critically evaluate the efficiency of the iPSC lentiviral transduction protocol used to introduce specific genes such as KLF4, c-MYC, OCT3/4, and SOX2 when generating IPSC or used to modify and genetically engineer iPSC lines. Assays verify true pluripotency by ensuring that the cells undergo trilineage differentiation into the three primary germ layers: ectoderm, mesoderm, and endoderm. Analyzing the gene expression and the resulting cells' functionality can indicate the differentiation potential of the iPSC line. When iPSC lines are generated from patients with known diseases, or a specific mutation is introduced to an IPSC line, genotyping can be performed to confirm the disease-associated mutations in the line.
Benefits of iPSC Assays
- Precision: Assays provide nuanced data about cell pluripotency and the presence of neurologically relevant genes.
- Consistency: Assays assure the quality of your cells and result in reproducible research outcomes, which are especially vital for complex neuronal studies.
- Validation: Assays guarantee the expression of pluripotency markers like KLF4, c-MYC, OCT3/4, and SOX2, or the presence of expected disease-associated mutations.
- Flexibility: Assays allow for the confirmation of unique reporters or the knock-down of specific genes in engineered iPSCs, ensuring customization of the cells for individual project needs.
- Safety: Assays confirm the absence of microorganism contamination (e.g., mycoplasm), karyotypic abnormalities, or any residual traces from the infection process (ie: SeV).
Understanding iPSC Assays
Technology
iPSC assays employ various technologies, from traditional microscopy for assessing iPSC morphology to advanced sequencing techniques for pluripotency and differentiation assays. Newer technologies, such as advanced fluorescence microscopy, have further expanded the scope of iPSC assays, increasing their ability to offer profound insights into pluripotency, cell identity, and cell growth.
iPSC Assay Services
Trilineage Differentiation
We’ve developed a unique transcription factor-based Trilineage Differentiation Assay protocol (also available as a kit) to validate differentiation potential into cells of all 3 germ layers
Lentiviral Transduction
Introducing genes into iPSC lines such as fluorescent reporters to visualize specific aspects of differentiation
Antibody Validation
Validating antibody specificity for immunocytochemistry using iPSCs
Flow Cytometry
Quantitative pluripotent marker expression assessment
Karyotyping
iPSCs sometimes carry karyotype abnormalities from source culture methods or from prolonged culture. This assessment is essential to ensure genetic integrity
RNA Sequencing
Gene expression profiling of iPSCs and iPSC-derived cells
Sendai Virus (SeV) Clearance
Assuring the purity of iPSCs post-generation
Sterility Testing
Ensuring the absence of microbial contamination
Immunocytochemistry
Visualizing specific characteristic markers in iPSCs (e.g., KLF4, c-MYC, OCT3/4, and SOX2) and iPSC-derived cells
Expansion & Cryopreservation
Generating and freezing down stocks of low-passage, high-quality iPSCs
Disease Genotype Confirmation
Verifying disease-specific markers, such as those for Parkinson’s or Alzheimer’s
iPSC Assay Techniques and Procedures
Techniques We Use
Standard Procedures
Elixirgen currently maintains iPSC cells using StemFit® Basic04 media and iMatrix-511 silk in a standard humidified 37°C, 5% CO2 incubator. Cells are passaged as single cells approximately once a week using EDTA/TrypLE™. Cell growth is closely monitored using a variety of methods including automated cell counting and observation of iPSC morphology. The protocols and reagents for StemFit® Basic04 and iMatrix-511 silk culture conditions are available from Elixirgen Scientific (Catalog Numbers: ASB04-C, NI511S).
Custom Procedures
At Elixirgen Scientific, we understand that every research project has unique requirements. That's why we're equipped to accommodate external protocols and assays allowing us to custom-tailor our approach to precisely fit the specifications of our customers. We have experience growing cells on a variety of substrates (including Matrigel®, Geltrex™, Laminin 521, and others) using a wide range of media (including mTeSR™, TeSR™-E8™, StemFlex™, and others). Although single-cell passaging offers many advantages, we are also adept at more traditional passaging methods requiring clumps of cells.
Range of Applications We Can Support
With our state-of-the-art technologies and methodologies, we support a myriad of research and clinical applications tailored to the needs of our partners and clients. Whether you're in early-stage research, drug discovery, or clinical therapeutics, our services are designed to propel your innovations forward.
Why Choose Elixirgen Scientific?
Expertise
With a team of highly skilled and experienced iPSC scientists and researchers, Elixirgen delivers accurate, reliable, and reproducible results.
Equipment and Resources
Elixirgen is equipped with cutting-edge technology, ensuring top-tier performance in iPSC assay services. Our location on the Johns Hopkins University Medical Campus provides us with a stimulating environment and access to additional resources.
Efficiency
Our expertise and resources enable us to deliver results with remarkable speed, without compromising on quality.
Scalability
Whether it's a small-scale academic project or a large-scale pharmaceutical study, Elixirgen can scale its services to meet your needs.
Cost-Effectiveness
By leveraging technology and optimizing processes, Elixirgen offers competitive pricing without sacrificing quality.
Frequently Asked Questions (FAQs)
Our iPSC assays are designed to ensure the quality and integrity of iPSCs. By confirming pluripotency, evaluating differentiation potential, and identifying any genetic abnormalities, our assays can provide the knowledge needed to proceed with your differentiations and achieve your research goals.
Absolutely! Elixirgen Scientific can customize assays based on your research requirements. Our team is experienced in developing and conducting a wide range of iPSC assays.
Elixirgen Scientific ensures quick turnaround times and high-quality results, allowing for seamless progression of your research. By outsourcing the iPSC expansion, differentiation, and assay steps, you can focus more on the actual research questions, thus increasing the efficiency of your work.
Absolutely. Elixirgen Scientific provides comprehensive support and guidance in the interpretation of assay results to help researchers better understand their iPSCs. We work closely with our clients to ensure they fully comprehend the data, thereby facilitating informed decision-making in their research process.
We pride ourselves on our collaborative approach. Our team can coordinate with your existing workflow, providing services at stages where you need support. With clear communication and efficient processes, we ensure our integration is smooth and beneficial to your research.
Our rigorous sterility testing protocol checks for microbial, fungal, and mycoplasma contamination, guaranteeing the purity of iPSCs.
Elixirgen offers competitive pricing without compromising on quality. Our expertise in iPSC culture ensures timely and precise results, making us a preferred choice for many in the field.
