Services

Cell-Based Assays

iFyber specializes in developing cell-based assays for safety and efficacy testing, offering a variety of standardized and custom functional bioassays to evaluate the effects of new biomaterials, medical devices, and investigational compounds on mammalian biological systems at the cellular and tissue levels.

Medical technologies employ a range of diverse functional materials, from synthetic polymers to recellularized ECM scaffolds. This diversity can lead to variable chemical and biological interactions at the cell or tissue interface. We implement high throughput, repeatable, relevant, and quantitative in vitro assays that provide clients with robust data that can be used to plan future product development and verification efforts.

Whether you are looking to transfer or optimize an existing assay for use with your technology or develop one from scratch, our cross-functional expertise in cell biology and interfacial chemistry ensures a swift path to completed assay development and reliable, actionable results.

Why Choose iFyber for Your Assay Needs:

Specialized Expertise and Investment in Your Research Success. iFyber focuses on specific research questions. This allows us to tailor our approach to your unique needs, ensuring the data we generate is directly relevant to your goals.

Standardized Assays with a Personalized Touch. While we offer standardized assays, we don't treat them as a simple "box to check." Our team of scientists is actively involved throughout the process, from assay selection and cell line considerations to data interpretation and scientific consultation.

Clear Communication and Collaboration. We believe in close collaboration with our clients, keeping you informed throughout the process and ensuring you fully understand the data and its implications for your research. High-touch, accessible support from a trusted team.

Connect with Our Scientists

Explore iFyber's Cell Culture & Tissue Services:

Biocompatibility and Cell/Biomaterial Interaction

Biocompatibility Assays:

Cytotoxicity Assays: Utilizing a range of cell lines/primary cells with methods including direct seeding, agar overlay/filter diffusion, or extracts. Employed assays are MTT, AlamarBlue, ATP, Flow cytometry, and others. Explore insights on the Pros and Cons of Cytotoxicity.
Skin Irritation Test: Utilizing a 3D in vitro model of human epidermal tissue.
Endotoxin/Pyrogenicity Assays: Performing LAL assay, MAT (Monocyte Activation Test), and rFC. Explore insights on ensuring patient safety through Endotoxin and Pyrogen Testing.
Other Available Biocompatibility Assays:
- Bright Field Microscopy
- Neutral Red Uptake
- Trypan Blue Exclusion
- Bacterial Reverse Mutation Assay
- Platelet activation
- Hemolysis assay

Cell/Biomaterial Interaction:

Stem Cell: Investigating interactions with biomaterials and assessing stem cell differentiation.
Immune Cells: Exploring M1/M2 polarization and monitoring cytokine release.
Osteoblasts: Evaluating osteoinductivity/osteoconductivity of biomaterials via ALP activity, Osteocalcin release, and Mineralization assay.
Endothelial Cells: Analyzing interactions with biomaterials and assessing vascularization potential.

Custom Assays (Representative Projects):

Electroporation Therapy Effects: Developing unique cell culture methods to evaluate the impact of a device on cancerous cells.
Collagen Coating Method: Pioneering a method to boost cell adhesion, enhancing biomaterial integration.

If your business requires a unique assay, we have the expertise to develop custom methods from the ground up, meeting your exact requirements.

Tissue Processing Efficiency

Decellulariziation Efficiency:

DNA Quantification: UVerification that the dsDNA content per mg of tissue/ECM does not surpass 50 ng.
Gel Electrophoresis: Validation that the length of residual DNA fragments is under 200 bp.
ECM Characterization: Ensuring the ECM maintains its native structural integrity.

Demineralization Efficiency:

Ca Quantification: Measurement of remaining calcium in DBM to ensure it does not exceed 8%.
BMP2 Quantification: Verification of BMP2 presence in DBM, crucial for its osteoinductive properties.

Recellularization Assay:

Cell infiltration: Seeding cells on decellularized tissues and evaluating their infiltration.
Degradation Profile: Monitoring the progressive changes in the presence of cells.

Custom Assays (Representative Projects):

Cellular Bone Matrix (CBM) Products: Evaluating cell viability of the product, determining the type of cells (stem cells, osteoblasts) present in their product, and evaluating osteogenicity.

If your business requires a unique assay, we have the expertise to develop custom methods from the ground up, meeting your exact requirements.

Wound Healing Evaluation

2D Cell Culture Assays:

Scratch Assay: Inducing a gap in the cell monolayer (epithelial cells, fibroblasts, Keloid fibroblasts) and tracking wound closure post-treatment to assess wound healing capabilities.
Transwell Cell Migration: Seeding cells on one side of transwell inserts and quantifying the migration to the opposite side of the membrane following exposure to the product, providing insights into cell motility and invasion.

3D In vitro Assay (HEE):

Human Epidermis model: Establishing a multilayer epidermal model to simulate epithelium, enabling evaluation of treatment efficacy in a more relevant context.

3D Ex vivo Assay:

Porcine Tissue: Establishing multispecies biofilms within porcine tissue samples to simulate complex wound environments, enabling evaluation of treatment efficacy in a more clinically relevant context.
Human Viable Skin Tissue: Inducing a burn wound infection in human skin tissue and evaluating the therapeutic effects of treatments on wound closure, bacterial count, and released cytokines.

Hemostasis Effects:

Thrombogenicity Testing: Assessment of relative hemoglobin absorbance (RHA) and the degree of thrombogenicity.

Advanced Cell & Tissue Analysis Techniques

Co-culture of Bacteria/Cells:

Macrophage/Gram-positive or Gram-Negative Bacteria: This technique is highly relevant for testing the efficacy of antimicrobial coatings or implants aimed at preventing infections. Additionally, it enables the investigation of immune cell activation and polarization in response to different materials, potentially in the presence of bacteria.

Co-culture of Virus/Cells:

Infectivity testing: Testing reagents to determine the parameters (i.e. time, ratio, etc.) required to inactivate a target virus. Testing a product or tissue provided by a company that needs confirmation of inactivation

Tissue Engineering:

Scaffolds Characterization: Physical & chemical characterization, biological evaluation, and degradation analysis of the engineered tissue constructs. Moreover, we leverage our extensive knowledge in tissue engineering to create customized co-culture systems tailored to specific research objectives.

Standarized Assays:

MTT Assay

What Is It?

The MTT assay provides a quantitative assessment of cell proliferation by measuring the activity of mitochondrial enzymes that reduce the tetrazolium salt, MTT, to formazan crystals that are solubilized with acidified isopropyl alcohol. The resulting purple solution is quantified spectrophotometrically at 570 nm. Standard intra-assay controls include vehicle-only (usually cell media) and a relevant mammalian cell toxin to demonstrate appropriate test system response.

Why Use It?

MTT is widely regarded as the go-to assay for in vitro cytotoxicity testing and is used regularly in industrial and academic R&D laboratories. The MTT assay has been demonstrated to be a relevant predictor of in vivo biocompatibility performance, highlighting its usefulness as a screening assay for new biomaterials or investigational compounds. Using ISO as a guideline, a material is considered cytotoxic if cell viability falls below 70% of untreated control cells.

Live/Dead Staining

What Is It?

The LIVE/DEAD assay is used to determine both cellular membrane integrity and cytosolic enzymatic function in response to a test article. Assay functionality is based on the ability of healthy cells to exclude ethidium homodimer-1, and at the same time cleave the membrane permeable, non-fluorescent calcein AM to calcein through functional cytosolic esterase activity, yielding green cytoplasmic fluorescence. Membrane-compromised cells are permeable to ethidium homodimer-1, which binds to nucleic acids resulting in nuclear-localized red fluorescence.

Why Use It?

Unlike most standard cytotoxicity assays, the LIVE/DEAD assay enables concurrent determination of two key aspects of cellular health: membrane permeability and enzymatic function. Healthy cells show green fluorescence, whereas dying or dead cells appear red. Quantification can be carried out by analyzing images that are captured using fluorescence microscopy methods or by using a fluorescence plate reader for high throughput purposes. As such, it provides more detail to the client and can help guide further focused testing.

Tunel Assay

What Is It?

Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) is used to identify DNA fragmentation, a terminal hallmark of both apoptotic and necrotic cells. Modified dUTPs are incorporated by the enzyme TdT at the 3’-OH ends of fragmented DNA. The modified dUTPs can be fluorescently labeled allowing quantification of apoptotic/necrotic cell number. This assay utilizes a double staining approach where total DNA appears blue and fragmented DNA appears red.

Why Use It?

BRDU Assay

What Is It?

The BrdU assay is based on the fluorometric detection of the thymidine analog BrdU (bromodeoxyuridine), upon its incorporation into genomic DNA during cell proliferation. BrdU may then be detected using anti-BrdU antibodies, allowing assessment of the population of cells that are synthesizing DNA. This indicates cell proliferation rate and a measurement of the number of cells in S phase of the cell cycle.

Why Use It?

If a test article demonstrates cytotoxicity as determined by MTT or NRU assays, the client may want to know more. One of the first aspects of cellular function to be investigated is DNA replication. The BrdU assay enables direct comparison between the proliferative status of cells treated with the test article and control cells. A reduction in BrdU uptake by treated cells indicates that DNA replication, and thus cell proliferation, is being negatively impacted.

Ready to elevate your research? Discover precision, expertise, and collaboration at iFyber. Contact us now to discuss your cell-based assay development needs.

Let's Discuss Your Needs