One of the benefits of cryopreserving cells is the flexibility in scheduling for later use that freezing provides. Determining the optimal solution for cryopreservation that consistently results in post-thaw recovery of the greatest number of highly viable, functional cells is crucial. The article by Sara S. Parker et al. published in eNeuro and recently added to BioLife’s Evidence library explains the process her group used for cryopreservation reagent selection and cell evaluation; applied to primary mouse embryonic cortical neurons for research.
Parker, S. S., Moutal, A., Cai, S., Chandrasekaran, S., Roman, M. R., Koshy, A. A., Khanna, R., Zinsmaier, K. E. and Mouneimne, G. (2018). High Fidelity Cryopreservation and Recovery of Primary Rodent Cortical Neurons. Eneuro; 5(5), ENEURO.0135-18.2018. doi:10.1523/eneuro.0135-18.2018
The authors compared cryopreservation of dissociated primary neurons in commercial cryopreservation media (including CryoStor® CS10 and CS5) and a home-brew solution composed of 10% DMSO in 50% serum/40% cell culture media. The optimal cryopreservation solution was determined by freezing neurons in each solution, storage in the vapor phase of liquid nitrogen, followed by assaying cells immediately post-thaw using trypan blue staining or 3 days post-thaw using calcein AM/ethidium homodimer-1 labeling. Readout was cell viability and viable yield post-thaw, compared to non-frozen neurons.
Following determination of the optimal storage solution, functionality of neural cells stored in CS10 was determined by assaying key neurodevelopmental markers, determining outgrowth and arborization of neuronal processes, and cells’ ability to establish functional neural networks.
Similarly, the team also demonstrated cryopreservation of primary embryonic mouse neurons in CS10 could be extended to include primary embryonic hippocampal neurons frozen in CS10.
The next article by Richard Harrison et al. published in Stem Cell Research & Therapy and recently added to BioLife’s Evidence library applies to storage and transport of cells intended for therapeutic use that have been labeled with magnetic particles.
Harrison, R., Lugo Leija, H. A., Strohbuecker, S., Crutchley, J., Marsh, S., Denning, C., El Haj, A. and Sottile, V. Development and validation of broad-spectrum magnetic particle labelling processes for cell therapy manufacturing. Stem Cell Research & Therapy. 2018; 9(1). doi:10.1186/s13287-018-0968-0
Separately, human bone marrow-derived mesenchymal stromal cells (MSCs), human neural stem cells (ReNs) and human iPS-derived cardiomyocytes (CMCs) grown in suspension were cultured with fluorescent labeled magnetic particles. Cell cultures were assessed for viability, senescence, and cell cycle status to determine that cell integrity was maintained following labeling. Labelling efficiency was determined and cell function was assessed by metabolic activity.
A key component in cell therapy manufacturing is the cells’ ability to be stored and transported. Magnetically labeled MSCs, ReNs and CMCs were stored 24 hours at 4°C in HypoThermosol®FRS.
Also recently added to BioLife’s Evidence library, Juliette Peltzer et al. in their review article published in frontiers in Immmunology, discuss MSC-based cell therapy for the treatment of Systemic Sclerosis.
Peltzer, J., Aletti, M., Frescaline, N., Busson, E., Lataillade, J. and Martinaud, C. Mesenchymal Stromal Cells Based Therapy in Systemic Sclerosis: Rational and Challenges. frontiers In Immunology. 2018; 9. doi:10.3389/fimmu.2018.02013
Regarding holding of MSCs at 4°C, the authors note, “To enhance cell stability, the final product can be stored at 4°C in a specialized hypothermic storage media such as HypoThermosol prior to injection…”
Which stage along the pathway to cell therapy commercialization are your cells at today? Biopreservation solutions play key roles at every stage. Plan ahead! Ask the Scientists. Consult our staff of cryopreservation experts for help optimizing your cell preservation processes. Call 1.866.6543 (toll-free in North America) or direct: 1.425.402.1400, today!
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