XR OrgCryo - Protein-Free Organoid Cryopreservation Solution

Description

Reliable organoid banking can be difficult because organoids are multicellular three-dimensional structures rather than uniform single-cell suspensions. Inadequate freezing conditions may lead to cryoinjury, structural disruption, delayed post-thaw recovery, or reduced expansion after organoids are returned to culture. These variables can affect experimental scheduling, preservation of valuable organoid lines, and reproducibility between studies.

XR Protein-Free Cryopreservation for Organoids is a ready-to-use organoid cryopreservation medium developed for the freezing and recovery of human and mouse organoids. The formulation supplies cryoprotective components and nutrients intended to reduce stress during freezing, cryogenic storage, and thawing while supporting recovery of organoid morphology and continued culture after thawing.

This protein-free organoid freezing medium contains 10% DMSO and is manufactured using cell culture-grade, pharmacopeial components. It contains no added human- or animal-derived ingredients and no protein components, helping researchers reduce exposure to undefined xenogeneic materials during organoid banking workflows.

With low endotoxin specifications, sterile filtration, and compatibility with controlled-rate freezing workflows, the medium is suitable for research laboratories establishing organoid cryobanks, preserving experimental organoid batches, or recovering organoids for subsequent expansion and downstream research.

Specifications

Features

• Specialized organoid cryopreservation medium for human and mouse organoid freezing workflows

• Protein-free formulation with no added human- or animal-derived components

• Contains 10% (v/v) DMSO to provide cryoprotective support during freezing

• Designed to reduce freeze-thaw stress and support post-thaw organoid recovery

• Suitable for short-term storage at −80°C followed by transfer to liquid nitrogen for long-term cryostorage

• Supports preservation of organoid morphology and continued expansion after recovery

• Ready-to-use format simplifies routine organoid banking and cryopreservation workflows

• Manufactured using cell culture-grade, pharmacopeial components

• Low-endotoxin formulation with an endotoxin specification of ≤0.1 EU/mL

• Filtered through a <0.2 μm filtration process

Performance Data

Liver Organoid Recovery Following Cryopreservation

Liver organoids were evaluated after freezing and thawing to observe post-thaw morphology and subsequent culture recovery. Representative microscopy images were collected immediately after recovery, after 24 hours, after 72 hours, and 72 hours following passage.

Representative liver organoid morphology immediately after thawing, during 72-hour recovery, and following passage.

Under the tested conditions, organoid viability after thawing was maintained above 80%. The recovered organoids progressively regained typical morphology during culture and could be used for downstream experiments after approximately one to two passages.

Applications

XR Protein-Free Cryopreservation for Organoids is intended for research workflows requiring organized storage, recovery, and continued culture of human or mouse organoids. The animal component-free and protein-free formulation is particularly useful where researchers need to reduce exposure to undefined biological materials during organoid banking.

• Organoid cryopreservation: Freezing cultured organoids for later recovery, expansion, characterization, or experimental use

• Organoid banking and biobanking: Establishing working or research organoid banks to preserve valuable organoid lines and reduce repeated tissue processing

• Human organoid cryopreservation: Preserving established human organoid cultures for scheduled experiments and long-term research storage

• Mouse organoid cryopreservation: Banking mouse-derived organoids used in developmental biology, disease modeling, or compound-response research

• Liver organoid cryopreservation: Freezing and recovering liver organoids while supporting post-thaw morphology and continued expansion

• Intestinal and colon organoid banking: Preservation of gastrointestinal organoid cultures after validation of an appropriate fragment size and recovery workflow

• Gastric, pancreatic, lung, and kidney organoid research: Cryostorage of organoid models after application-specific workflow optimization

• 3D organoid culture workflows: Coordinating organoid production, cryogenic storage, and recovery across multi-stage experimental projects

• Disease-model research: Maintaining cryopreserved organoid batches for repeat testing, method validation, and comparative studies

• Compound screening studies: Recovering banked organoids to support experiments requiring comparable starting material across multiple runs

• Organoid quality-control workflows: Preserving reference or backup cultures for morphology, growth, viability, and marker-analysis studies

• Long-term organoid storage: Controlled-rate freezing followed by transfer to liquid nitrogen for extended cryogenic preservation

Recommended Cryopreservation Workflow

1. Recover Organoids from the Culture Matrix

At the end of the culture period, mechanically detach the matrix droplets from the culture plate and transfer them to a centrifuge tube. Pipette repeatedly to help separate the organoids from the matrix.

2. Pellet the Organoids

Centrifuge at 2500 rpm for 3–4 minutes, then carefully remove the supernatant and residual matrix without disturbing the organoid pellet.

3. Resuspend in Cryopreservation Medium

Slowly add the organoid freezing medium and gently resuspend the organoid pellet. Avoid excessive pipetting that may cause unwanted mechanical damage. The supplied protocol uses organoids collected from approximately two wells of a 24-well plate for one cryovial as a general reference.

4. Perform Controlled-Rate Freezing

Place the cryovials in a controlled-rate freezing container and transfer them to a −80°C freezer. Storage at −80°C is intended for short-term preservation. For long-term organoid storage, transfer the cryovials to liquid nitrogen after approximately 12 hours at −80°C.

5. Thaw and Recover the Organoids

Rapidly thaw the vial using a 37°C water bath or a suitable cell-thawing device. Transfer the organoids and freezing medium to a centrifuge tube and add at least three volumes of organoid culture medium.

6. Remove the Cryopreservation Medium

Centrifuge to remove the DMSO-containing freezing medium. Resuspend the recovered organoids in an appropriate volume of extracellular matrix according to organoid number and the laboratory's validated culture method.

7. Resume Organoid Culture

For a 24-well plate workflow, the supplied protocol uses approximately 50 μL of matrix per well. Allow the matrix to solidify at 37°C for approximately 20 minutes, then add the appropriate organoid culture medium and continue incubation.

Why Use a Protein-Free Organoid Freezing Medium?

Traditional cell freezing media may contain serum, albumin, or other biologically derived components that introduce undefined variables into sensitive organoid workflows. These components can vary between lots and may be undesirable for laboratories seeking a more controlled organoid cryopreservation process.

XR Protein-Free Cryopreservation for Organoids provides a protein-free and animal component-free alternative for organoid banking. The formulation helps reduce exposure to xenogeneic biological materials while retaining 10% DMSO for cryoprotection during freezing.

The ready-to-use format also reduces the need to prepare freezing mixtures immediately before use, helping laboratories standardize organoid freezing procedures across operators, experimental batches, and organoid research projects.

FAQ

For Research Use Only. Not intended for diagnostic, therapeutic, or clinical applications.