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CellMatrix Basement Membrane Gel

ACS-3035


Volume: 5 mL
Storage: -80°C

CellMatrix Basement Membrane Gel is a soluble, growth factor reduced basement membrane extract that does not contain phenol red. CellMatrix Gel is purified from murine Engelbreth-Holm-Swarm (EHS) tumor. Major components include laminin, collagen IV, entactin and heparin sulfate proteoglycan.


Product category
Reagents
Comments
Each lot of CellMatrix Gel is shown to promote the attachment of hESCs/hiPSCs and to maintain hESCs/hiPSCs in a pluripotent state as determined by the expression of Oct-4 and Nanog pluripotent markers.
Store at -80°C in aliquots. Avoid repeated freezing and thawing.
Applications
Stem cell research
Cell growth and viability
Volume
5.0 mL
Product format
Frozen
Storage conditions
-80°C or colder
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Price: $221.00 EA
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Documentation

Biosafety Icon BSL 1

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Detailed product information

General

Specific applications
  • Three-dimensional culture of primary tissue derived human and mouse organoids
  • Three-dimensional culture of iPSC derived organoids
  • Three-dimensional culture of tumorspheres from various cell lines
  • Sandwich culture of human hepatocytes
  • Feeder-free culture of human embryonic stem cells (hESC) and induced pluripotent stem cells (hiPSC)
  • Culture of iPSCs-derived neural progenitor cells (NPCs) and neurons
  • Angiogenesis modeling via the tube formation assay

Handling information

Handling procedure

Preparation and storage

Critical: CellMatrix Membrane Gel must remain frozen at all times during storage. Once thawed, keep CellMatrix on ice at all times. Do not allow it to warm above 4°C or premature gelation will occur.

Procedure

  1. Thaw stock CellMatrix Membrane Gel vial on ice in a 4°C refrigerator overnight.
  2. Determine the appropriate volume per aliquot based on concentration and application. Please refer to the certificate of analysis for the lot­specific concentration.
  3. Dispense appropriately­sized aliquots using pre­cooled tips into pre­cooled tubes on ice and re-freeze immediately.
  4. Thaw aliquots on ice. Once thawed, do not re-freeze. Once thawed, use within 7 days.
A. Three-dimensional culture of organoids in CellMatrix
  1. Thaw an aliquot of CellMatrix on ice and keep on ice.
  2. Resuspend dissociated organoids (fragments or single cells) in undiluted CellMatrix. Do introduce bubbles when pipetting. Work quickly to ensure the suspension remains cold.
  3. Dispense as small droplets in a pre-warmed (in a 37°C cell culture incubator) tissue culture treated vessel (e.g., 50 µL per well in a 24-well plate). Work quickly to prevent premature gelation.
  4. Place the plate in a 37°C cell culture incubator for 15-30 minutes to induce gelation.
  5. Add an appropriate volume of organoid culture media.

Harvesting organoids from CellMatrix

  1. Aspirate the media from the well completely
  2. Using a cell lifter scrape the well surface to release the gelled domes
  3. Transfer the domes to a 15 conical tube.
  4. Pipet up and down 20-30 times to break up the domes and release the organoids from the CellMatrix gel.
  5. Wash the organoids in cold DMEM:F12 (ATCC 30-2006) by centrifugation.
  6. Aspirate the supernatant leaving behind the pellet of organoids.
  7. Proceed with downstream assays using the organoids or resuspend in fresh CellMatrix to continue culture.
B. Sandwich culture of human hepatocytes
  1. Thaw hepatocytes and seed on collagen type-1 coated plates.
  2. After 6-24 hours, perform a complete media change with cold hepatocyte maintenance media that has been supplemented with 0.25 mg/mL CellMatrix. Refer to the certificate of analysis for the lot­specific concentration. For example, if the lot-specific concentration of CellMatrix is 15 mg/mL, add 16 uL CellMatrix per mL of hepatocyte culture media.
  3. Return plate to the incubator.
  4. After 24 hours perform a complete media change with hepatocyte maintenance media (not supplemented with CellMatrix).
C. Angiogenesis modeling via the tube formation assay
  1. Thaw CellMatrix Gel in the refrigerator (2°C to 8°C), in ice, overnight.
  2. Mix well by carefully pipetting CellMatrix Gel up and down, being careful not to introduce air bubbles.
  3. Do not dilute. Pipette 150 µL/cm2 onto the tissue culture vessel surface.  
  4. Swirl the vessel gently to ensure that the entire surface is evenly covered.
  5. Leave the coated dishes at 37°C for 30 minutes.
  6. The coated vessels are ready for use. 

D. Feeder-free culture of human embryonic stem cells (hESC) and induced pluripotent stem cells (hiPSC)

Refer to the ATCC Stem Cell Culture Guide for more information.

Important: CellMatrix Gel will solidify in 15 to 30 minutes if the temperature is above 15°C. Keep CellMatrix Gel and labware (pipette tips, serological pipettes, conical tubes) on ice at all times to prevent the matrix from gelling prematurely. If air bubbles form when coating the dishes, use a chilled pipette tip to break up the bubbles.

For stem cells, NPCs, NPCs-derived neurons:

  1. Thaw CellMatrix Gel in the refrigerator (2°C to 8°C), in ice, overnight.
  2. Dilute the thawed CellMatrix Gel to 150 µg/mL by directly adding CellMatrix in cold DMEM: F­12 Medium (ATCC 30­-2006) on ice and mix well. Immediately coat each 6 cm dish with 2 mL diluted CellMatrix Gel.
  3. Swirl dish gently to ensure that the entire dish is evenly covered.
  4. Leave the coated dishes at 37°C for one hour.
  5. Aspirate the coating solution and immediately plate the cells. It is critical that the coating does not dry out.
  6. If the dishes will not be used the same day they are prepared, do not aspirate the coating solution. Seal the coated dishes with parafilm and store at 2°C to 8°C for up to one week. Note that stored dishes should be warmed to room temperature in a biological safety cabinet for at least one hour before use.

References

  1. ATCC Angiogenesis Technical Bulletin.
  2. Albini A, et al. A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res 47(12): 3239­-3245, 1987.
  3. Kubota Y, et al.  Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J Cell Biol 107:1589-­1598, 1988.
  4. Ponce M, et al. Identification of endothelial cell binding sites on the laminin gamma 1 chain. Circ Res 84:688­-694, 1999.
  5. Eisenstein M, Thinking outside the dish. Nat Methods 3:1035­-1043, 2006.
  6. Arnaoutova I, et al. The endothelial cell tube formation assay on basement membrane turns 20: state of the science and the art. Angiogenesis 12(3):267-­274, 2009.
  7. Angel M, Yanik MF. Innate immune suppression enables frequent transfection with RNA encoding reprogramming proteins. PLoS ONE. 5(7):e11756, 2010.

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Endotoxin
< 8 EU/mL by LAL assay
Functional tests
Each lot of CellMatrix Gel is shown to promote the attachment of hESCs/hiPSCs and to maintain hESCs/hiPSCs in a pluripotent state as determined by the expression of Oct-4 and Nanog pluripotent markers.

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Bergqvist C, et al. An inner nuclear membrane protein induces rapid differentiation of human induced pluripotent stem cells. Stem Cell Res 23:33-38, 2017. PubMed: 28668644

Frequently Asked Questions

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