The CliniMACS Prodigy® Adherent Cell Culture process allows automation of midbrain dopaminergic (mDA) progenitor cell differentiation from pluripotent stem cells (PSCs), in a scalable and functionally closed system.
Cell therapies using PSC-derived mDA progenitor cells represent a novel and promising route for the treatment of Parkinson’s disease (PD). The CliniMACS Prodigy Adherent Cell Culture process offers nine modules that can automate various steps during the mDA progenitor cell differentiation process, such as inoculation and culture of starting PSCs, medium changes for neural cell induction and differentiation. These modules can be flexibly combined to effectively adapt established manual processes on the CliniMACS Prodigy. Meanwhile, the GMP compliant, single-use CliniMACS Prodigy Tubing Set 730 combined with external culture vessels provide a sterile, closed, and scalable environment for the PSC differentiation into mDA progenitor cells. In addition, if the resulting cells need to be further purified after differentiation, the MACSQuant® Tyto® Cell Sorter offers a gentle and GMP-compliant cell sorting solution.
The coating module of the CliniMACS Prodigy Adherent Cell Culture process can be used to coat the CliniMACS Prodigy chamber or external culture vessels with customized coating solutions prior to differentiation. Cryopreserved PSCs can be resuspended in the starting cell bag which then, using the inoculation module, can be inoculated in pre-coated culture vessels based on designed seeding density.
If differentiation takes place in the CliniMACS Prodigy chamber, the culture module allows users to set up cell culture conditions, such as temperature and CO2 concentration. If differentiation takes place in external culture vessels, an external incubator is needed to provide required cell culture conditions.
The efficient differentiation of mDA progenitor cells relies on high-quality recombinant cytokines. Our recombinant cytokines and growth factors are available in three quality grades: research grade, premium grade, and MACS GMP Grade, to facilitate a seamless transition from basic research to clinical application. Human SHH (C24II), the essential cytokine during mDA progenitor cell differentiation, is currently available in research and premium grades with MACS GMP grade coming soon.
Additionally, during mDA progenitor cell differentiation, the media change module can be used to apply various mediums to support neural cell induction, proliferation, and differentiation processes.
At the end of differentiation, the harvest module can be employed to dissociate the resulting mDA progenitor cells from the culture vessels into the CentriCult™ Unit for centrifugation. Subsequently, cells can be resuspended in the desired medium and harvested in a target cell bag for downstream applications.
Flow cytometric analyses are commonly used for quality control of PSC-derived mDA progenitor cells. The CliniMACS Prodigy Tubing Set 730’s integrated sampling pouches allow for safe separation and collection of control samples during the differentiation process. With the MACSQuant Analyzer family of flow cytometers and our REAfinity Recombinant Antibodies, Miltenyi Biotec provides standard solutions for flow cytometric analyses of PSC-derived mDA progenitor cells.
Complement to the MACSQuant Analyzers, we also offer Express Modes. These algorithm-based tools make use of thousands of data sets for processing, gating, and analysis of samples, allowing you to standardize and automate flow cytometry analyses. In addition, customized Express Modes can be designed based on specific needs and processes. The MACSQuant Instruments and the MACS Flow Cytometry portfolio are for research use only.
After differentiation, flow cytometric quality control analyses were applied using a MACSQuant Analyzer. The cells differentiated on the CliniMACS Prodigy developed a typical midbrain dopaminergic phenotype, comparable to the cells differentiated manually in 12-well plates at lab scale. The resulting cells exhibited high percentage of FoxA2 and OTX2 (mDA progenitor cell markers) double-positive cells, lacked expression of undesired neural lineage markers PAX-6 and Sox1, pluripotency marker Oct3/4, and showed a decreased proliferative capacity measured by Ki-67 expression.
After differentiation, if the resulting cells need to be further purified, the MACSQuant Tyto Cell Sorter offers a gentle and GMP-compliant cell sorting solution. In the PD trial led by Prof. Jun Takahashi at CiRA (Kyoto, Japan), the MACSQuant Tyto Cell Sorter was used for GMP-compliant sorting of dopaminergic progenitor cells derived from induced pluripotent stem (iPS) cells. The results revealed that cells sorted by the MACSQuant Tyto Cell Sorter showed higher post-survival rates and final yields, increased neural fiber outgrowth, and reduced sorting time, in comparison to other devices. Read more about this exciting story in the scientific poster below.
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