Cell Freezing and Preservation Media
Store Your Cells and Tissues with Optimized Cryopreservation Media
Storage and cryopreservation of cells and tissues are vital parts of the workflow for biological research. Your choice of freezing medium can influence the quality of your banked cells and future experiments. Ensure high cell viability and functional stability following storage and thawing by using a reliable cryopreservation medium offered by STEMCELL Technologies.
Choose from cGMP-manufactured, protein- and/or serum-free freezing and biopreservation media to preserve your cells and tissues. For long-term storage of your cells in low temperature environments (-80°C to -196°C), opt for a ready-to-use freezing medium such as CryoStor® CS10 or mFreSR™. For short-term hypothermic preservation (2 – 8°C), you can use a preservation medium such as HypoThermosol® FRS. Explore all our cell freezing and preservation media to meet your cryopreservation workflow needs for different cell types.
Cryopreservation Basics: Protocols and Best Practices for Freezing Cells
Learn the fundamentals of cell cryopreservation and find best practices for freezing down your cells in this resource.
cGMP-Manufactured Cell Freezing and Preservation Media
CryoStor® Freezing Media
Cell and Tissue Types:
- Immune cells, hepatocytes, human peripheral blood mononuclear cells (PBMCs), Chinese Hamster Ovary (CHO) cells, myeloma cell lines, hybridomas, human mesenchymal stromal cells (MSCs), human embryonic and induced pluripotent stem cells (ES and iPS cells), mouse organoids, and other extremely sensitive cell and tissue types.
Recommended For:
- Long-term storage of cells and tissues at low-temperature environments (-80°C to -196°C)
Features:
- Designed to mitigate temperature-induced molecular stress responses during freezing and thawing
Formulation:
- CryoStor® CS2, CryoStor® CS5, and CryoStor® CS10 are pre-formulated with 2%, 5%, and 10% USP-grade DMSO, respectively.
- CryoStor® CSB is the DMSO-free base formulation of CryoStor® products that can be used in your workflow to reduce the level of cryopreservation-induced delayed-onset cell death.
- cGMP-manufactured
BloodStor® Freezing Media
Cell Types:
- Cord blood and tissue, peripheral blood and bone marrow
Recommended For:
- Cryopreservation of stem cells and other cells isolated from umbilical cord blood, peripheral blood, bone marrow, and other biologics
Features:
- Formulated to be compatible with many automated stem cell banking systems
Formulation:
- BloodStor® 55-5 is preformulated with 55% (w/v) DMSO USP, 5% (w/v) Dextran-40 USP, and water-for-injection (WFI)-quality water
- BloodStor® 100 contains 100% (w/v) DMSO USP
- cGMP-manufactured
HypoThermosol® FRS Preservation Media
Cell Types:
- All cells and tissues including pluripotent stem cells, hematopoietic stem and progenitor cells, and mesenchymal stromal cells
Recommended For:
- Short-term storage and/or shipment of cells at 2 – 8°C rather than at cryogenic temperatures
Features:
- Uniquely formulated to address the molecular-biological response of cells during the hypothermic preservation process
Formulation:
- HypoThermosol® FRS preservation media is a serum-, protein-, and animal component-free medium
- cGMP-manufactured
Why Use cGMP-Manufactured Freezing Media?
Smooth your path through the regulatory landscape with cGMP-manufactured freezing and preservation media formulated using USP-grade ingredients. By using cGMP-manufactured freezing media, you can ensure that the products are consistently produced and controlled according to quality standards; an important consideration when you plan to move to the clinic in the future.
Defined and Serum-Free Cryopreservation Media
mFreSR™ and FreSR-S™ Cryopreservation Media
Cell Types:
- Human embryonic stem (ES) and induced pluripotent stem (iPS) cells
Recommended For:
- Cryopreservation of ES and iPS cells cultured in TeSR™ media
Features:
- Higher thawing efficiencies than with conventional methods using serum1-4
- FreSR™-S is optimized for cryopreservation of cells in single cell suspension
Formulation:
- FreSR™-S is an animal component-free (ACF) medium, optimized for cryopreservation of cells in single cell suspension
MesenCult™-ACF Cryopreservation Medium
Cell Types:
- Mesenchymal stem cells (MSCs)
Recommended For:
- Cryopreservation of MSCs, including human MSCs previously cultured in MesenCult™-XF or MesenCult™-ACF media
Features:
- Thawed MSCs have reproducibly high recovery rates and maintain MSC multipotency and expansion capacities
Formulation:
- MesenCult™-ACF Freezing Medium is a defined, serum-free and animal component-free medium that contains DMSO
STEMdiff™ Neural Progenitor Freezing Medium
Cell Types:
- Neural progenitor cells (NPCs) derived from human pluripotent stem cells
Recommended For:
- Cryopreservation of NPCs generated from ES or iPS cells using STEMdiff™ Neural Induction Medium
Features:
- NPCs can be frozen at any point post-neural induction with reproducibly high recovery rates
- Post-thaw, NPCs display healthy morphology, express NPC markers and retain the potential to expand and differentiate into neurons, astrocytes and other neural cell types
Formulation:
- STEMdiff™ Neural Progenitor Freezing Medium as a defined and serum-free medium containing DMSO
Why Use Serum-Free Cryopreservation Media?
Lab-made formulations of freezing medium often contain fetal bovine serum (FBS). However, FBS contains undefined components, such as growth factors and hormones, and its use in freezing media raises concerns about lot-to-lot variability and the risk of transmitting potentially infectious agents. FBS is usually not recommended for cell banking, commercial, or clinical applications, such as the production of biologicals.
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.
Figure 3. mFreSR™ Improves Thawing Efficiencies 5- to 10-Fold over Other Reported Methods
H9 hESCs were cryopreserved in mFreSR™ at the indicated passage number. Thawing efficiencies were analyzed by counting the number of surviving clumps after thawing.
Related Products to Complete Your Freezing & Thawing Workflow
Complete your cryopreservation workflow by preserving your cells and tissues in sterile, self-standing cryogenic vials of different sizes and cap colors. Or obtain consistent freezing profiles using an alcohol-free container ideal for the cryopreservation of most cells and cell lines. Explore products for sterile cryostorage and freezing below:
For downstream processing, researchers can thaw their cryopreserved cells using the ThawSTAR® CFT2 Automated Thawing System—a water-free instrument that delivers cell thawing profiles similar to those of a water bath. Conveniently thaw your cells in the biosafety cabinet, in ~2.5 minutes, while you prepare for the next step in your experiment.
Standardize Your Cell Thawing Process
Increase confidence in your cell thawing workflow by ensuring sample sterility and consistent thawing performance with ThawSTAR® CFT2—a sensor-based automated thawing system.
Step-by-Step Cell Thawing Demonstration
Follow this step-by-step protocol video to see how ThawSTAR® CFT2 could fit in your laboratory’s cell thawing workflow.
Protocols and Technical Tips for Freezing and Thawing Cells
Cryopreservation and Thawing of ES/iPS Cells
Cryopreservation and thawing of human ES or iPS cells using either mFreSR™, CryoStor® CS10, or FreSR™-S.
How to Thaw Frozen Primary Cells
Step-by-step protocol for thawing frozen primary cells.
References
- Fujioka T, et al. (2004) A simple and efficient cryopreservation method for primate embryonic stem cells. Int J Dev Biol 48(10): 1149-54
- Ha SY, et al. (2005) Cryopreservation of human embryonic stem cells without the use of a programmable freezer. Hum Reprod 20(7): 1779-85
- Ji L, et al. (2004) Cryopreservation of adherent human embryonic stem cells. Biotechnol Bioeng 88(3): 299-312
- Ware CB, et al. (2005) Controlled-rate freezing of human ES cells. Biotechniques 38(6): 879-80, 882-3
