SCIENCE
Isolated Mesenchymal
Stem Cell (MSC) Exosomes
Optimal-Purity Exosomes for Clinical Research
Mesenchymal Stem Cells
Unlocking Regenerative Potential
Harnessing the healing power of mesenchymal stem cell exosomes begins with isolating young, healthy mesenchymal stem cells (MSCs).
MSCs and other progenitor cells reside in many different types of tissue. The tissue type, in which MSCs reside, has a significant impact on the behavior of these progenitor cells. For example, neural stem cells may produce exosomes that have a favorable profile for regenerating damaged neural tissue, and cancer stem cells produce exosomes that promote the development of tumors. MSCs residing in the chorionic plate of the placenta are the youngest precursor cells in the connective tissue lineage. The natural purpose of the placenta is to support rapid, controlled development of fetal tissues, while modulating the natural immune and inflammatory response and suppressing malignant degeneration.
Screening
Exceeding The Highest Standards
In order to create a safe, isolated, placental MSC line, Kimera Labs® has implemented a strict screening process that exceeds the standards required for tissue banks and cGMP manufacturers.
Similar to the screening process performed by tissue banks, donors are tested for HIV, HTLV, Hepatitis B, Hepatitis C, Syphilis and CMV. Kimera Labs® goes beyond these FDA requirements by testing for these viruses and additional testing for Zika, EBV, Hepatitis E, HHV6, HHV7, HHV8, HSV1, HTLV1, HTLV2, BKV, and Norovirus. Exosomes produced by Kimera Labs® remain under quarantine prior to release for research use until testing confirms that they are negative for these pathogens.
In addition to understanding the rigorous screening and testing process for pathogens, it is important to note that placental mesenchymal stem cells are not a natural reservoir for these viruses. Any possible risk of contamination of donor tissues despite rigorous screening is furtherer reduced during the MSC culture and exosome isolation process. Finally, sterility and endotoxin assays are performed after the final stage of production to confirm the absence of any infectious agents.
Isolation
Sterile Preparation of Purified MSC Exosomes
At every step of its proprietary MSC exosome production process, Kimera Labs strives to achieve the highest purity of stem cells, culture media and exosomes.
Using ISO 7 clean rooms and ISO 5 isolation hoods, the Kimera Labs® quality measures begin before the exosome isolation process begins. Fresh cultures of isolated, neonatal MSCs are initiated regularly using culture medium that contains no animal or human products. Unlike many laboratories that use fetal bovine serum or human serum to more easily culture cells, Kimera Labs uses xeno-free chemically defined medium
to reduce the potential for any type of contamination. As the MSCs release exosomes into the culture media, this conditioned media is sterile-filtered to remove the cells. After separating the MSC exosomes from the cells, the conditioned media undergoes a proprietary, serial purification process that yields isolated MSC exosomes in scalable quantities.
Characterization
Defining Exosome Detection Methods
Kimera Labs® has pioneered innovative techniques and technologies to more precisely characterize and quantify exosomes for quality assurance and exploration of exosome-based diagnostic modalities.
Generally, exosome characterization has utilized the most widely available types of biologic content assays to define properties such as protein concentrations, and more recently, particle counts. Nano Tracking Analysis (NTA), which uses light scattering from nanoparticles to provide discrete particle quantification and size distribution, is commonly used to validate the concentration of exosomes in research samples. NTA provides an accurate measure of exosomes in a sample only if the sample consists of pure exosomes. Any other nanoparticulate material, such as viruses or antibody-antigen complexes that may be present in a biologic sample, can cause a. falsely elevated assessment of the number of exosomes in that sample.
Because NTA cannot distinguish between nanoparticulate debris and exosomes, other exosome detection modalities are necessary to confirm the nature and number of exosomes in a sample. The substantial error that can occur with counting exosomes along with other nanoparticles in amniotic fluid or processed tissue, such as bone marrow or placental tissue, can not only result in significantly exaggerated counts of the number of exosomes in a sample, but also means that there may be a large amount of nanoparticulate debris remaining. Kimera Labs® uses much more precise exosome detection modalities such as dSTORM microscopy to ensure the MSC origin and quality of its exosome products.
Identification
High-Quality, Isolated MSC Exosomes
Kimera Labs® leads the way in defining the character of exosomes, in order to ensure the purity and consistency of its pharmaceutical-grade exosomes.
Beyond the basic modalities of protein analysis, flow cytomery, RNA sequencing and Nano Tracking Analysis, Kimera Labs uses advanced imaging modalities to define the character of its exosome products for use in development of clinical applications. Kimera Labs has visualized its pure MSC exosome products with electron microscopy and atomic force microscopy to verify that the particles in solution are exosomes based on their surface morphology. Now, using STORM super resolution microscopy, Kimera Labs is able to visualize individual proteins characteristic of MSC exosomes such as CD 9, 63 and 81 on the surface of its exosomes. Not only can this technique accurately count exosomes,
it can directly verify that they are in fact MSC exosomes. Beyond characterizing Kimera exosomes, STORM super resolution microscopy can actually visualize internalization of exosomes into live cells and track their movement within the cell. Kimera Labs uses this advanced imaging technology to verify each batch of its pharmaceutical-grade exosomes for quality and consistency.
Research & Development
Advancements in Exosome Technology
Using advanced knowledge of exosome identification and characterization techniques, Kimera Labs scientists and physicians are developing new clinical diagnostic modalities based on circulating exosomes.
Kimera Labs currently utilizes multi-modality exosome characterization technologies such as ELISA protein analysis, high pressure liquid chromatography, RNA sequencing and NanoSight Nano Tracking Analysis, atomic force microscopy, electron microscopy and STORM super resolution fluorescence microscopy. These technologies are now being used to develop the next generation of diagnostic modalities. Knowing that the microRNA content and membrane composition of exosomes is distinctly different in different disease states and with aging, Kimera Labs has taken the first steps toward defining exosome profiles that may be used as reference standards for future clinical diagnostic tests. Exosome-based diagnostics would provide a superior method of more accurate, early detection and evaluation of disease severity. Performing a ‘liquid biopsy’ using exosomes readily accessible in body fluids could avoid the morbidity of surgical biopsy and may help to diagnosis life-threatening diseases like cancer at much earlier stages, where treatment is much more successful.