CryoStor® CS10

Animal component-free, defined cryopreservation medium with 10% DMSO
From: 245 USD

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Animal component-free, defined cryopreservation medium with 10% DMSO
From: 245 USD

Overview

CryoStor® CS10 is a uniquely formulated serum-free, animal component-free, and defined cryopreservation medium containing 10% dimethyl sulfoxide (DMSO). Designed to preserve cells in low temperature environments (-80°C to -196°C), CryoStor® CS10 provides a safe, protective environment for cells and tissues during the freezing, storage, and thawing processes. CryoStor® CS10 is recommended for the cryopreservation of hepatocytes, tissue samples, human peripheral blood, CHO cells, myeloma cell lines, hybridomas, human mesenchymal stem cells, human embryonic and human induced pluripotent stem cells (ES cells and iPS cells) and other extremely sensitive cell types. CryoStor® CS10 is cGMP-manufactured with USP grade components.
Advantages:
• Ready-to-use
• Serum-free, protein-free
• Animal component-free
• cGMP manufactured with USP grade / highest-quality components
• FDA master file
• Sterility, endotoxin, and cell-based quality control testing
Contains:
• 10% dimethyl sulfoxide (DMSO)
• Other ingredients
Cell Type:
B Cells; CHO Cells; Hematopoietic Stem and Progenitor Cells; Hybridomas; Intestinal Cells; Macrophages; Mesenchymal Stem and Progenitor Cells; Monocytes; Myeloma; NK Cells; Other; Pluripotent Stem Cells; T Cells
Species:
Human; Mouse; Rat; Non-Human Primate; Other
Brand:
CryoStor
Area of Interest:
Cord Blood Banking; Epithelial Cell Biology; Immunology; Stem Cell Biology
Formulation:
Animal Component-Free; Serum-Free; Defined

Scientific Resources

Educational Materials

(7)

Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

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Workflow Stages

Data and Publications

Data

Figure 1. Immune Cells Cryopreserved in CryoStor®CS10 Show Reproducibly High Post-Thaw Cell Viability

CryoStor®CS10 effectively mitigates temperature-induced molecular cell stress responses to maximize post-thaw viability and recovery for a variety of immune cell types, including T cells (data not shown) and B cells. Here, human B cells from 6 different donors cryopreserved in CryoStor®CS10 show reproducibly high viability after thawing, as measured by Propidium Iodide staining (ranging from 94.3 - 97.9%).



Figure 2. Immune Cells Cryopreserved in CryoStor®CS10 Retain Functionality Post-Thaw

(A) Human peripheral blood Pan-T cells cryopreserved in CryoStor®CS10 were thawed and cultured with or without the addition of T cell activating factors. Cells from Donors 1-3 were cultured in RPMI Medium supplemented with 10% FBS, with (activated) or without (control) 40 ng/mL PMA and 1 ug/mL Ionomycin for 24 hours. Cells from Donors 4-5 were cultured in ImmunoCult™-XF T Cell Expansion Medium (Catalog #10981), with (activated) or without (control) ImmunoCult™ Human CD3/CD28 T Cell Activator (Catalog #10971) for 48 hours. Supernatants were collected from the cultures, and concentrations of secreted cytokines were determined using the Human IL-2 ELISA Kit (Catalog #02006). Activation by either PMA and Ionomycin or ImmunoCult™ Human CD3/CD28 T Cell Activator led to increased secretion of IL-2 compared to unstimulated control cultures. (B) Human B cells (Donors 6 - 11) cryopreserved in CryoStor®CS10 were thawed and activated with 1 µg/mL CD40 and 100 ng/mL IL-21 for 7 days. Supernatants were collected from the cultures and immunoglobulin G (IgG) production was measured using the Human IgG ELISA Antibody Pair Kit (Catalog #01994). Compared to unstimulated control cultures, B cell activation led to increased IgG​ ​secretion.

Publications

(42)
Nature methods 2017 APR

Marker-free coselection for CRISPR-driven genome editing in human cells.

Agudelo D et al.

Abstract

Targeted genome editing enables the creation of bona fide cellular models for biological research and may be applied to human cell-based therapies. Therefore, broadly applicable and versatile methods for increasing its efficacy in cell populations are highly desirable. We designed a simple and robust coselection strategy for enrichment of cells with either nuclease-driven nonhomologous end joining (NHEJ) or homology-directed repair (HDR) events by harnessing the multiplexing capabilities of CRISPR-Cas9 and Cpf1 systems. Selection for dominant alleles of the ubiquitous sodium/potassium pump (Na(+)/K(+) ATPase) that rendered cells resistant to ouabain was used to enrich for custom genetic modifications at another unlinked locus of interest, thereby effectively increasing the recovery of engineered cells. The process is readily adaptable to transformed and primary cells, including hematopoietic stem and progenitor cells. The use of universal CRISPR reagents and a commercially available small-molecule inhibitor streamlines the incorporation of marker-free genetic changes in human cells.
Cell cycle (Georgetown, Tex.) 2016 JAN

Non-integrating episomal plasmid-based reprogramming of human amniotic fluid stem cells into induced pluripotent stem cells in chemically defined conditions.

Slamecka J et al.

Abstract

Amniotic fluid stem cells (AFSC) represent an attractive potential cell source for fetal and pediatric cell-based therapies. However, upgrading them to pluripotency confers refractoriness toward senescence, higher proliferation rate and unlimited differentiation potential. AFSC were observed to rapidly and efficiently reacquire pluripotency which together with their easy recovery makes them an attractive cell source for reprogramming. The reprogramming process as well as the resulting iPSC epigenome could potentially benefit from the unspecialized nature of AFSC. iPSC derived from AFSC also have potential in disease modeling, such as Down syndrome or $\$-thalassemia. Previous experiments involving AFSC reprogramming have largely relied on integrative vector transgene delivery and undefined serum-containing, feeder-dependent culture. Here, we describe non-integrative oriP/EBNA-1 episomal plasmid-based reprogramming of AFSC into iPSC and culture in fully chemically defined xeno-free conditions represented by vitronectin coating and E8 medium, a system that we found uniquely suited for this purpose. The derived AF-iPSC lines uniformly expressed a set of pluripotency markers Oct3/4, Nanog, Sox2, SSEA-1, SSEA-4, TRA-1-60, TRA-1-81 in a pattern typical for human primed PSC. Additionally, the cells formed teratomas, and were deemed pluripotent by PluriTest, a global expression microarray-based in-silico pluripotency assay. However, we found that the PluriTest scores were borderline, indicating a unique pluripotent signature in the defined condition. In the light of potential future clinical translation of iPSC technology, non-integrating reprogramming and chemically defined culture are more acceptable.
Oncotarget 2016 FEB

Successful expansion of functional and stable regulatory T cells for immunotherapy in liver transplantation.

Safinia N et al.

Abstract

Strategies to prevent organ transplant rejection whilst minimizing long-term immunosuppression are currently under intense investigation with regulatory T cells (Tregs) nearing clinical application. The clinical trial, ThRIL, recently commenced at King's College London, proposes to use Treg cell therapy to induce tolerance in liver transplant recipients, the success of which has the potential to revolutionize the management of these patients and enable a future of drug-free transplants. This is the first report of the manufacture of clinical grade Tregs from prospective liver transplant recipients via a CliniMACS-based GMP isolation technique and expanded using anti-CD3/CD28 beads, IL-2 and rapamycin. We report the enrichment of a pure, stable population of Tregs (textgreater95% CD4(+)CD25(+)FOXP3(+)), reaching adequate numbers for their clinical application. Our protocol proved successful in, influencing the expansion of superior functional Tregs, as compared to freshly isolated cells, whilst also preventing their conversion to Th17 cells under pro-inflammatory conditions. We conclude with the manufacture of the final Treg product in the clinical research facility (CRF), a prerequisite for the clinical application of these cells. The data presented in this manuscript together with the much-anticipated clinical results from ThRIL, will undoubtedly inform the improved management of the liver transplant recipient.
Stem Cell Reviews and Reports 2016 AUG

Functionalizing Ascl1 with Novel Intracellular Protein Delivery Technology for Promoting Neuronal Differentiation of Human Induced Pluripotent Stem Cells

Robinson M et al.

Abstract

Pluripotent stem cells can become any cell type found in the body. Accordingly, one of the major challenges when working with pluripotent stem cells is producing a highly homogenous population of differentiated cells, which can then be used for downstream applications such as cell therapies or drug screening. The transcription factor Ascl1 plays a key role in neural development and previous work has shown that Ascl1 overexpression using viral vectors can reprogram fibroblasts directly into neurons. Here we report on how a recombinant version of the Ascl1 protein functionalized with intracellular protein delivery technology (Ascl1-IPTD) can be used to rapidly differentiate human induced pluripotent stem cells (hiPSCs) into neurons. We first evaluated a range of Ascl1-IPTD concentrations to determine the most effective amount for generating neurons from hiPSCs cultured in serum free media. Next, we looked at the frequency of Ascl1-IPTD supplementation in the media on differentiation and found that one time supplementation is sufficient enough to trigger the neural differentiation process. Ascl1-IPTD was efficiently taken up by the hiPSCs and enabled rapid differentiation into TUJ1-positive and NeuN-positive populations with neuronal morphology after 8 days. After 12 days of culture, hiPSC-derived neurons produced by Ascl1-IPTD treatment exhibited greater neurite length and higher numbers of branch points compared to neurons derived using a standard neural progenitor differentiation protocol. This work validates Ascl1-IPTD as a powerful tool for engineering neural tissue from pluripotent stem cells.
Vox sanguinis 2016 AUG

Relevance of flow cytometric enumeration of post-thaw leucocytes: influence of temperature during cell staining on viable cell recovery.

Fritsch G et al.

Abstract

BACKGROUND AND OBJECTIVES Our post-thaw cell recovery rates differed substantially in interlaboratory comparisons of identical samples, potentially due to different temperatures during cell staining. MATERIALS AND METHODS Viable CD34(+) cells and leucocyte (WBC) subtypes were quantified by multiparameter single-platform flow cytometry in leucapheresis products collected from 30 adult lymphoma and myeloma patients, and from 10 paediatric patients. After thawing, cells were prepared for analysis within 30 min between thawing and acquisition, at either 4°C or at room temperature. RESULTS For cell products cryopreserved in conventional freezing medium (10% final DMSO), viable cell recovery was clearly lower after staining at 4°C than at RT. Of all WBC subtypes analysed, CD4(+) T cells showed the lowest median recovery of 4% (4°C) vs. 25% (RT), followed by CD3, CD34 and CD8 cells. The recovery was highest for CD3γδ cells with 44% (4°C) vs. 71% (RT). In the 10 samples cryopreserved in synthetic freezing medium (5% final DMSO), median recovery rates were 89% for viable CD34 (both at 4°C and RT) and 79% (4°C) vs 68% (RT) for WBC. CONCLUSIONS The post-thaw environment and, potentially, the cryoprotectant impact the outcome of cell enumeration, and results from the analysis tube may not be representative of the cells infused into a patient.
THIS PRODUCT IS MANUFACTURED UNDER A cGMP QUALITY MANAGEMENT SYSTEM COMPLIANT TO 21 CFR 820.
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