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Cryostor 10

Manufactured by BioLife Solutions
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CryoStor 10 is a serum-free, protein-free, and chemically defined cryopreservation medium. It is designed to maintain the viability and functionality of cells during freezing and thawing processes.

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Lab products found in correlation

Sodium heparin vacutainer tube, by BD (1 mentions) Ficoll pacque, by GE Healthcare (1 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions) Vacutainer cpt tube, by BD (1 mentions) Cryomed controlled rate freezer, by Thermo Fisher Scientific (1 mentions) Stem cellbanker, by AMS Biotechnology (1 mentions) D2650, by Thermo Fisher Scientific (1 mentions)

8 protocols using cryostor 10

1

QCE System Product Cryopreservation

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The products of the QCE system runs were frozen in Cryostor10 (Biolife Solutions, Bothell, WA) and stored at −120°C in lN2. Representative vials for each run were release tested as described below to ensure cells met all relevant release criteria for a clinical cell bank.
Tirughana R., Metz M.Z., Li Z., Hall C., Hsu D., Beltzer J., Annala A.J., Oganesyan D., Gutova M, & Aboody K.S. (2018). GMP Production and Scale-Up of Adherent Neural Stem Cells with a Quantum Cell Expansion System. Molecular Therapy. Methods & Clinical Development, 10, 48-56.
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2

PBMC Isolation from Venous Blood

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Approximately 60 mL of venous blood was collected in sodium heparin vacutainer tubes (BD Biosciences, catalog no. 367874) and shipped overnight from the clinical site to OncoSec Medical at ambient temperature. Heparin tubes were immediately processed for peripheral blood mononuclear cells (PBMC) using standard Ficoll-Pacque (GE) gradient extraction as described previously (1 (link)). PBMCs were frozen in CryoStor 10 (BioLife Solutions) and stored in liquid nitrogen until use.
Algazi A.P., Twitty C.G., Tsai K.K., Le M., Pierce R., Browning E., Hermiz R., Canton D.A., Bannavong D., Oglesby A., Francisco M., Fong L., Pittet M.J., Arlauckas S.P., Garris C., Levine L.P., Bifulco C., Ballesteros-Merino C., Bhatia S., Gargosky S., Andtbacka R.H., Fox B.A., Rosenblum M.D, & Daud A.I. (2020). Phase II Trial of IL-12 Plasmid Transfection and PD-1 Blockade in Immunologically Quiescent Melanoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(12), 2827-2837.
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3

Cryogenic Banking of Clinical-Grade CD-NSCs

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To produce the current GMP (cGMP) clinical bank of CD-NSCs, eight frozen vials of clinical-grade CD-NSCs (frozen at passage 26) were thawed at 37°C using an established SOP (SOP-1955A). Cell number was determined using trypan blue staining, and cells were diluted to 5.2 × 107 cells/100 mL NSC medium, transferred into eight cell inlet bags, and expanded in the QCE system according to SOP-1955A. Harvested cells were pooled and frozen at 6 × 107cells/2.5 mL Cryostor10 (Biolife Solutions, Bothell, WA). Cells were aliquoted (2.5 mL per vial) into 4-mL externally threaded polypropylene cryogenic vials with a conical bottom (Corning 430662) and frozen in a controlled rate freezer (Planer PLC, Middlesex, UK) at cooling rates of −0.1°C/min to 10°C/min until cells reached −80°C, at which point they were transferred to the vapor phase of a liquid nitrogen tank. In total, 260 vials were frozen and stored.
Tirughana R., Metz M.Z., Li Z., Hall C., Hsu D., Beltzer J., Annala A.J., Oganesyan D., Gutova M, & Aboody K.S. (2018). GMP Production and Scale-Up of Adherent Neural Stem Cells with a Quantum Cell Expansion System. Molecular Therapy. Methods & Clinical Development, 10, 48-56.
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4

Cryopreservation of hiPSC Aggregates

Cited 1 time
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A solution of non-DMSO cryoprotective agents (CPAs) was formulated using sucrose, glycerol, L-isoleucine, poloxamer 188 (P188), and human serum albumin as previously optimized for the cryopreservation of hiPSC aggregates (Li et al., 2020 (link)). A commercially available DMSO-based CPA solution (CryoStor 10, BioLife Solutions) was used in comparison. Neuronal cells were incubated in the CPA solution at room temperature. Viable and nonviable cell counts were based on membrane integrity using 10 µM acridine orange (AO) and 15 µM propidium iodide (PI) and measured at 0, 30, and 60 min.
Li R., Walsh P., Truong V., Petersen A., Dutton J.R, & Hubel A. (2021). Differentiation of Human iPS Cells Into Sensory Neurons Exhibits Developmental Stage-Specific Cryopreservation Challenges. Frontiers in Cell and Developmental Biology, 9, 796960.
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5

PBMC Isolation and Cryopreservation

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Approximately 60 mL of venous blood was collected in BD Vacutainer CPT tubes (BD Biosciences, catalog no. 367874) and spun at 1,500 to 1,800 RCF for at least 15 minutes within 2 hours of blood collection. Spun tubes were then shipped overnight from the clinical site to OncoSec Medical at ambient temperature. The top portion of the tubes was removed and washed twice in DPBS (Gibco) and PBMCs frozen in CryoStor 10 (BioLife Solutions) and stored in liquid nitrogen until use.
Algazi A.P., Twitty C.G., Tsai K.K., Le M., Pierce R., Browning E., Hermiz R., Canton D.A., Bannavong D., Oglesby A., Francisco M., Fong L., Pittet M.J., Arlauckas S.P., Garris C., Levine L.P., Bifulco C., Ballesteros-Merino C., Bhatia S., Gargosky S., Andtbacka R.H., Fox B.A., Rosenblum M.D, & Daud A.I. (2020). Phase II Trial of IL-12 Plasmid Transfection and PD-1 Blockade in Immunologically Quiescent Melanoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(12), 2827-2837.
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6

Cryopreservation of Bioengineered Renal Epithelial Cells

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SLA-BRECS or IM-BRECS were flushed and incubated with room temperature HTS-Purge Solution (# 637112 BioLife Solutions, Inc., Bothell, WA) for 2 minutes. HTS-Purge was then replaced with cryopreservation buffer, CryoStor 10 (CS10, #640222 BioLife Solutions) at 4°C, and the inlet and outlet were capped with sterile, Nylon, male luer integral lock ring plugs (Value Plastics, Fort Collins, CO) to prepare HREC BRECS for cryostorage. A CryoMed controlled rate freezer (Thermo Scientific, Waltham, MA) was utilized to freeze complete, individual SLA-BRECS or IM-BRECS. Briefly, this protocol consisted of a cooling rate of −1°C/min sample temperature to −4°C, followed by a rapid cooling phase during phase change, −25°C/min chamber temperature to −40°C. The chamber was then warmed 10°C/min to −14°C. After phase change, a cooling rate of −1°C/min sample temperature to −40°C was used. The last rapid cooling step was −10°C/min sample temperature to −90°C. BRECS were then directly transferred into the gas phase of a liquid nitrogen tank for cryostorage.
Pino C.J., Westover A.J., Buffington D.A, & Humes H.D. (2017). Bioengineered Renal Cell Therapy Device for Clinical Translation. ASAIO journal (American Society for Artificial Internal Organs : 1992), 63(3), 305-315.
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7

Cryopreservation of Cultured Cells

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Flasks were washed with pre-warmed 37°C PBS without Ca 2+ and Mg 2+ and incubated for 15 min at 37°C, 5% CO 2 /21% O 2 with TrypLE Select. TrypLE Select was deactivated using DTI, and cells were scraped from the flask surface using a cell scraper (VWR, 734-2604). After centrifugation at 300g for 3 min, cells were resuspended in NSÀ/À with P/S and filtered with a 40-mm cell strainer before counting. Cells were frozen at a standard cell density of 5 £ 10 5 À5 £ 10 6 cells/vial according to the manufacturing instructions. Cells were again centrifuged and resuspended in an adequate volume in cryopreservation solutions (STEM-CELLBANKER [11897; Amsbio, Abingdon, UK], CryoStor 10 [210374; BioLife Solutions, Bothell, WA, USA] and 10% DSMO [D2650; Thermo Fisher Scientific]) in NSÀ/À and transferred to a cryotube, placed in a CoolCell that allows gradual temperature reduction, frozen in a À80°C freezer for a day and then transferred to a nitrogen tank.
Baqué-Vidal L., Main H., Petrus-Reurer S., Lederer A.R., Beri N.E., Bär F., Metzger H., Zhao C., Efstathopoulos P., Saietz S., Wrona A., Jaberi E., Willenbrock H., Reilly H., Hedenskog M., Moussaud-Lamodière E., Kvanta A., Villaescusa J.C., La Manno G, & Lanner F. (2024). Clinically compliant cryopreservation of differentiated retinal pigment epithelial cells. Cytotherapy, 26(4).
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8

Adipose-Derived Stem Cell Injection for Submandibular Glands

Cited 2 times
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Eligible patients were randomized in a 1:1 ratio to receive ASCs or placebo. Placebo consisted of CryoStor10 (BiolifeSolutions), the freeze media for ASCs, containing 10% dimethyl sulfoxide (DMSO). The ASCs were provided by the Cardiology Stem Cell Centre (CSCC) - Rigshospitalet. ASCs were isolated from abdominal adipose tissue from three healthy female donors, according to good manufacturing practice (GMP) in clean room facilities as described previously (16, 17 (link)). Only one donor's cells were used in each cell vial. A total of 0.5 mL of ASCs or placebo was injected without anaesthesia into each submandibular gland, and for the patients receiving ASCs, this corresponded to a dose of approximately 25 × 106 cells per gland. The injections were performed ultrasound-guided by a trained investigator, as described previously (15 (link)).
Jakobsen K.K., Carlander A.L., Todsen T., Melchiors J., Paaske N., Østergaard Madsen A.K., Kloch Bendtsen S., Mordhorst C., Stampe H., Kastrup J., Ekblond A., Haack-Sørensen M., Farhadi M., Maare C., Friborg J., Lynggaard C.D., Werner Hauge A., Christensen R., Grønhøj C, & von Buchwald C. (2024). Mesenchymal Stem/Stromal Cell Therapy for Radiation-Induced Xerostomia in Previous Head and Neck Cancer Patients: A Phase II Randomized, Placebo-Controlled Trial. Clinical Cancer Research, 30(10), 2078-2084.
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