BioTether Sciences - A boutique CRO focused on bioanalysis and cell-based assays located in North Bay

Boutique CROs emerge as the preferred choice for clients seeking personalized attention, specialized expertise, flexibility, direct communication, and cost-effective solutions in the biotech industry. Their nimble approach and client-centric focus make them invaluable partners for biopharmaceutical companies looking to achieve their research and development goals with precision and efficiency.

Whether you're a startup, emerging biotech company, or well-established pharmaceutical firm, partnering with a small CRO can help you:

• ➡️ Accelerate your path to success
• ➡️ Streamline your research processes
• ➡️ Bring your groundbreaking therapies to market faster.

Join BioTether Sciences for an engaging webinar discussing strategies and techniques

 to study the pharmacokinetics, bio-distribution, pharmacodynamics, immunogenicity, and the potency

of cell, gene, and protein therapeutics.

https://attendee.gotowebinar.com/recording/7058460459861473536

Primary human dermal fibroblasts are extremely helpful to support research and development of drugs and devices. Material obtained from the skin biopsy can be utilized in several investigative procedures such as examination of cellular morphology, immunofluorescence staining of biochemical markers,  enzymatic studies, and molecular biology. A fibroblast is a specific type of connective tissue cell that is found in skin. In genetics research, cultured fibroblasts are particularly important because they can be isolated and grown from a small biopsy from forearm skin cells. The cells can be examined for genotypes and grown and tested for disease associated phenotypes. Human dermal fibroblasts are useful to study extracellular matrix production and wound healing. They can also be used as model systems to study many aspects of cell physiology, reprogramming, and induced pluripotency. Among the various uses of a skin biopsy, the most frequent is the confirmation and testing for clinical diagnoses. More recently primary human skin fibroblasts are used to study CAS9-CRISPR mediated gene editing and knock-out. Gene therapies using lentivirus, AAV, and nanoparticles can be delivered to primary human fibroblasts.

In the specific area of inborn errors of metabolism, cell culture from skin biopsies are used for enzymatic measurements to provide a diagnosis. For example, primary fibroblasts are used for the definitive diagnosis and study of Mucopolysaccharidoses. A hallmark of these lysosomal storage diseases is the accumulation of glycosaminoglycans and deficiencies in the enzymes that process them. Many other inherited metabolic disorders can be studied using primary dermal fibroblasts.  Similarly, other genetic diseases affecting the skin, muscle, connective tissue, bone, and immune system may be diagnosed and studied using skin fibroblasts. Disorders such as Parkinson’s, cancer, heart disease may be studied using primary fibroblasts or cells derived from them.  

One exciting aspect of working with these cells is the ability to dedifferentiation fibroblasts to an artificial stem cell type by the induction of pluripotent cells (iPSCs). This can be accomplished by the forced expression of certain factors. The iPSCs can be then directed toward other cell lineages. The multipotent plasticity of these cells and the  ability to isolate them from a consenting individual patient for autologous treatment opens many avenues for research and therapeutic development.

Primary human dermal fibroblasts may be collected with little discomfort by taking a small (1-8mm) forearm skin biopsy punch. The aseptically collected tissue is then transported in cell culture media containing serum and antibiotics by overnight courier. Once in our laboratory, BioTether Sciences can culture, study, and preserve these cells. We have the cell culture and bioanalytical expertise to support your program and advance the research and development of your therapeutic (www.BioTether.com).

Mycoplasma are an unusual type of bacteria that can infect animal, insect, and plant cells. They can be a very costly pest for biopharmaceutical researchers and manufacturers. These little organisms (0.2-2 micron) are the smallest and simplest free-living parasite. They can impact the safety and quality of cell-derived biopharmaceutical products. This puts patients at risk and could cause untimely and costly delays to manufacturing. To minimize these risks, routine testing for mycoplasma should be performed throughout the product research, development and manufacturing process.

There are several ways to test for mycoplasma.  These include direct culture, Hoechst DNA staining, PCR-based testing, and  hybridization-amplification by ELISA. For the production of biopharmaceuticals, direct culture-based approaches assess the presence of viable cells using indicator cell lines exposed to mycoplasma. However, this type of testing requires weeks and costs approximately $5000 per sample. An alternative approach to identifying mycoplasma contamination is through PCR-based or hybridization ELISA testing. The molecular-based PCR method or the hybridization-ELISA method is ideal for research laboratories. These methods are rapid, highly sensitive, specific, reliable, and cost-effective. The PCR-based method and hybridization method are designed to amplify or bind to the conserved 16S rRNA region of the mycoplasma genome. Eight species of mycoplasma are responsible for approximately 95% of cell culture contamination events and are detected using these techniques. There are numerous PCR based protocols (for example see ATCC.org kit offering) that use primers recognizing the 16S rRNA and can detect mycoplasma contamination by measuring a distinct PCR product. For the hybridization technique, (i.e. MycoProbe, Mycoplasma detection kit by R&D Systems) samples are hybridized in a microplate containing biotin labeled capture oligonucleotide probes and digoxigenin-labeled detection probes that map to the 16S rRNA sequence. The hybridization product is detected with anti-digoxigenin alkaline phosphatase.  PCR and hybridization techniques have been demonstrated to be extremely sensitive, comparable to direct culture, are rapid, and cost significantly less than direct culture (approximately $500 per sample).

Enhanced regulated testing with an overlay of quality assurance systems to comply with GMP requirements adds to the cost. For GMP lot release, Europe and the USA-FDA have established guidances to follow. For research and development, PCR and hybridization-ELISA techniques are ideal for routine, rapid screening. Cell lines can be tested every few months or screened immediately after receipt or before/after cryopreservation, therefore, allowing cell lines and biological products to be used after a short quarantine. BioTether Sciences performs both PCR based, and hybridization-ELISA based mycoplasma testing to support your research and development. This service is included free with cell based assay and stable cell line development projects placed at BioTether Sciences. We can also provide rapid turn-around and reporting on client supplied samples (cell line supernatant, cell pellets, reagents). Don’t let those little parasites derail your biopharmaceutical program!

Reference: Drexler HG, Uphoff CC. Mycoplasma contamination of cell cultures: Incidence, sources, effects, detection, elimination, prevention. Cytotechnology 39: 75-90, 2002. PMCID: PMC3463982