ABOUT MSC’S
EXOSOMES DERIVED FROM ISOLATED MESENCHYMAL STEM CELLS (MSCS)
High-Purity Exosomes for Clinical Investigation
Mesenchymal Stem Cells: Unleashing Regenerative Capacity
Initiating the utilization of mesenchymal stem cell exosomes’ therapeutic potential starts with the isolation of youthful, robust mesenchymal stem cells (MSCs).
MSCs, alongside various progenitor cells, inhabit diverse tissue types, each influencing the behavior of these precursor cells significantly. For instance, neural stem cells may generate exosomes with advantageous attributes for regenerating impaired neural tissue, while exosomes from cancer stem cells may stimulate tumor growth. Among these, MSCs residing in the chorionic plate of the placenta represent the most juvenile precursor cells within the connective tissue lineage. The placenta naturally orchestrates the swift, controlled development of fetal tissues while modulating immune responses, inflammation, and suppressing malignant transformations.
SCREENING
Meeting and Surpassing Stringent Standards
To ensure the creation of a secure and isolated placental MSC line, Innovare Biologics® has implemented a stringent screening process that surpasses the standards mandated for tissue banks and cGMP manufacturers.
Similar to the protocol conducted by tissue banks, donors undergo testing for HIV, HTLV, Hepatitis B, Hepatitis C, Syphilis, and CMV. Innovare Biologics ® exceeds these FDA requirements by conducting additional testing for Zika, EBV, Hepatitis E, HHV6, HHV7, HHV8, HSV1, HTLV1, HTLV2, BKV, and Norovirus. Exosomes produced by Innovare Biologics ® remain under quarantine before release for research purposes until testing confirms their negativity for these pathogens.
It’s important to note that placental mesenchymal stem cells are not a natural reservoir for these viruses, further minimizing the risk of contamination despite rigorous screening. Additionally, during the MSC culture and exosome isolation process, any potential contamination is further reduced. Lastly, sterility and endotoxin assays are conducted after the final production stage to ensure the absence of any infectious agents.
ISOLATION
Ensuring Sterile Preparation of Purified MSC Exosomes
Throughout its proprietary MSC exosome production process, Innovare Biologics® consistently pursues the utmost purity of stem cells, culture media, and exosomes. The quality measures employed by Innovare Biologics® commence even before the exosome isolation process, utilizing ISO 7 clean rooms and ISO 5 isolation hoods. Fresh cultures of isolated neonatal MSCs are routinely initiated using culture medium devoid of any animal or human products. Unlike numerous laboratories that rely on fetal bovine serum or human serum for cell culture, Innovare Biologics® opts for xeno-free chemically defined medium, minimizing the risk of contamination. As MSCs release exosomes into the culture media, this conditioned media undergoes sterile filtration to eliminate cells. Following the separation of MSC exosomes from cells, the conditioned media undergoes a proprietary, serial purification process, yielding isolated MSC exosomes in scalable quantities.
Characterization
Establishing Methods for Exosome Detection
Innovare Biologics® has spearheaded pioneering techniques and technologies aimed at meticulously characterizing and quantifying exosomes to ensure quality assurance and advance exosome-based diagnostic methods.
Traditionally, exosome characterization has relied on widely available biologic content assays to determine properties such as protein concentrations and, more recently, particle counts. Nano Tracking Analysis (NTA), which leverages light scattering from nanoparticles to offer discrete particle quantification and size distribution, is commonly employed to validate exosome concentration in research samples. However, NTA provides accurate measurements only in samples containing pure exosomes. Presence of other nanoparticulate materials, such as viruses or antibody-antigen complexes, in a biologic sample can lead to falsely elevated assessments of exosome numbers.
Given NTA’s inability to differentiate between nanoparticulate debris and exosomes, additional exosome detection modalities are essential to confirm the nature and quantity of exosomes in a sample. The significant margin of error inherent in counting exosomes alongside other nanoparticles in amniotic fluid or processed tissue—like bone marrow or placental tissue—not only leads to potentially exaggerated counts of exosome numbers but also implies a substantial amount of residual nanoparticulate debris. Innovare Biologics® employs highly precise exosome detection modalities such as dSTORM microscopy to ensure the MSC origin and quality of its exosome products.
IDENTIFICATION
Isolation of High-Quality MSC Exosomes
Innovare Biologics® leads the forefront in defining the composition of exosomes, aiming to ensure the purity and uniformity of its pharmaceutical-grade exosomes.
In addition to conventional techniques such as protein analysis, flow cytometry, RNA sequencing, and Nano Tracking Analysis, Innovare Biologics® employs advanced imaging modalities to characterize its exosome products for clinical application development. The lab has utilized electron microscopy and atomic force microscopy to visualize its pure MSC exosome products, confirming their identity based on surface morphology. Presently, leveraging STORM super-resolution microscopy, Innovare Biologics® can visualize individual proteins characteristic of MSC exosomes, such as CD 9, 63, and 81, on the exosome surface. This technique not only enables accurate exosome counting but also directly confirms their identity as MSC exosomes.
Beyond characterization, STORM super-resolution microscopy facilitates the visualization of exosome internalization into live cells and tracks their movement within the cell. Innovare Biologics® leverages this cutting-edge imaging technology to validate every batch of its pharmaceutical-grade exosomes for quality and uniformity.
RESEARCH & DEVELOPMENT
Progress in Exosome Technology
Leveraging advanced exosome identification and characterization techniques, scientists and physicians at Innovare Biologics® are pioneering the development of novel clinical diagnostic methods based on circulating exosomes.
Currently, Innovare Biologics® employs a range of multi-modality exosome characterization technologies, including ELISA protein analysis, high-pressure liquid chromatography, RNA sequencing, NanoSight Nano Tracking Analysis, atomic force microscopy, electron microscopy, and STORM super-resolution fluorescence microscopy. These cutting-edge technologies are being harnessed to create the next generation of diagnostic modalities.
Recognizing that the microRNA content and membrane composition of exosomes vary significantly across different disease states and with aging, Innovare Biologics® has embarked on the initial stages of establishing exosome profiles that could serve as reference standards for future clinical diagnostic tests. Exosome-based diagnostics offer the potential for superior, more precise detection and assessment of disease severity.
By conducting a “liquid biopsy” utilizing exosomes readily found in bodily fluids, this approach could circumvent the morbidity associated with surgical biopsies and potentially enable the diagnosis of life-threatening diseases like cancer at much earlier stages, greatly improving treatment outcomes.
