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View additional product information for Geltrex™ hESC-Qualified, Ready-To-Use, Reduced Growth Factor Basement Membrane Matrix - FAQs (A1569601)
18 product FAQs found
No, the Ready-To-Use Geltrex matrix was developed for use with the StemPro hESC SFM System. It is not recommended for other PSC media systems.
For other PSC media systems, we recommend the Geltrex LDEV-Free, hESC-Qualified, Reduced Growth Factor Basement Membrane Matrix (Cat. Nos. A1413301, A1413302).
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Geltrex hESC-Qualified, Ready-To-Use, Reduced Growth Factor Basement Membrane Matrix (Cat. No. A1569601) is a pre-diluted Geltrex product ready to use for coating culture vessels. There is no need to dilute the solution further. The product has also been QC tested using hESCs to ensure that every lot will maintain the attached pluripotency of these cells. The full product description can be found here:
Geltrex hESC-Qualified, Ready-To-Use, Reduced Growth Factor Basement Membrane Matrix (Cat. No. A1569601)
Geltrex LDEV-Free, hESC-Qualified, Reduced Growth Factor Basement Membrane Matrix (Cat. Nos. A1413301, A1413302) is a "concentrated" version of the Geltrex hESC-Qualified, Ready-To-Use above. You can pre-dilute this concentrated solution to pre-coat the culture vessel to grow hESCs at your desired usage concentration. This Geltrex matrix has also been QC tested using hESCs to ensure every lot will maintain the attached pluripotency of these cells. The full product description can be found here:
Geltrex LDEV-Free, hESC-Qualified, Reduced Growth Factor Basement Membrane Matrix (Cat. Nos. A1413301, A1413302)
Geltrex LDEV-Free Reduced Growth Factor Basement Membrane Matrix (Cat. Nos. A1413201, A1413202) is the same as the product above. However, it has not been tested to verify it will support hESC growth and expansion. This is a "general" version and not specific to a cell type or line. A full product description can be found here:
Geltrex LDEV-Free Reduced Growth Factor Basement Membrane Matrix (Cat. Nos. A1413201, A1413202)
Geltrex LDEV-Free Reduced Growth Factor Basement Membrane Matrix (Cat. Nos. A1413201, A1413202)
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The Gentamycin sulfate concentration in Geltrex hESC-Qualified, Ready-To-Use, Reduced Growth Factor Basement Membrane Matrix (Cat. No. A1569601) is 10 µg/mL.
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For stem cells, a 1:100 dilution is the recommended starting dilution for Geltrex hESC, Ready-To-Use, Reduced Growth Factor Basement Membrane Matrix (Cat. No. A1569601). Other (lower) dilutions may be necessary for other primary cell types and should be determined empirically.
Geltrex hESC-Qualified, Ready-To-Use, Reduced Growth Factor Basement Membrane Matrix is in principle a diluted version of the frozen Geltrex LDEV-Free, hESC-Qualified, Reduced Growth Factor Basement Membrane Matrix.
Please take a look at page 2 under section “Recover frozen PSCs in complete StemFlex Medium” in the manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0016431_StemFlexMediumKit_UG.pdf) for the volume of Geltrex to use to cover a 6-well, 12-well, or 24-well plate.
Basically,
One well of a 24-well plate has a surface area of 2 cm2 and you need 300 µL of diluted Geltrex per well.
One well of a 48-well plate has a surface area of 1 cm2 and you need 150 µL of diluted Geltrex per well.
One well of a 96-well plate has a surface area of 0.32 cm2 and you need 48 µL (˜50 µL) of diluted Geltrex per well.
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We do not have comparison data for the two Geltrex products. We recommend using the Geltrex LDEV-Free, hESC-Qualified, Reduced Growth Factor Basement Membrane Matrix for hepatocytes. We have not performed a significant amount of testing using the Geltrex hESC-Qualified, Ready-To-Use, Reduced Growth Factor Basement Membrane Matrix on these cells.
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Human ES cells are generally characterized by their typical morphology (they grow as tightly packed clusters of small cells with high ratio of nucleus to cytoplasm); surface marker expression; RT-PCR detection of stem cell-specific gene expression (such as Oct3/4, Sox2, and Nanog); alkaline phosphatase staining, and telomerase activity assay. The most commonly used ES specific surface markers include stage-specific embryonic antigens SSEA-3 and SSEA-4 for human ES cells. Other ES-specific surface antigens also include TRA-1-60 and TRA-1-81. (Science 282:1145 (1998).
Human ES cells are derived from human blastocyst inner cell masses, isolated by immunosurgery with rabbit antiserum to BeWO cells (a human trophoblast cell line) (Science 282:1145 (1998)).
Embryonic stem (ES) cells are derived from the early mammalian embryo and are capable of unlimited, undifferentiated proliferation in vitro while maintaining their potential to differentiate into a wide of range of adult tissues including germ cells. The pluripotency of the ES cells is normally demonstrated in vitro by inducing ES cells to differentiate into embryoid bodies and checking lineage-specific markers for differentiated cells in three body layers (endo, meso, and ectoderm), or injecting them into immunodeficient mice and determining the cell types produced in the teratomas.
Yes, this is normal. It has been observed in product development. Remove the Geltrex solution after pre-coating and add cells as normal. H9 human embryonic stem cells have been cultured on dishes pre-coated with Ready-to-Use Geltrex Matrix with this observation in the pre-coating procedure.
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Yes, try a variety of dilutions to coat the plates. Some customers use a 1:100 dilution and have better results. The optimal dilution is cell-line dependent.
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Geltrex Matrix is a basement membrane extract that contains collagen IV, laminin, and other matrix proteins and growth factors that are required for cell adhesion. AlgiMatrix Matrix contains only alginate which does not facilitate cell adhesion. There is nothing in the alginate sponges for the cells to interact with.
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Ready-to-Use Gibco Geltrex Matrix provides only a coating for cell attachment and was developed mainly for ESC and iPSC use. Because the protein concentration is not high enough, it should not be used for thick gel or 3D applications.
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Yes, this is normal. It has been observed in product development. Remove the Gibco Geltrex Matrix solution after pre-coating and add cells as normal. H9 human embryonic stem cells have been cultured on dishes pre-coated with Ready-to-Use Gibco Geltrex Matrix with this observation in the pre-coating procedure.
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If you would like to culture the cells once recovered from the matrix, we recommend washing the cells first, then using trypsin/EDTA, TrypLE reagent, or a solution of 3,000 U/mL collagenase and 1,000 U hyaluronidase in low-glucose DMEM. If you would like to monitor cell surface markers, we recommend removal of the cells by mechanical means, such as a vigorous wash or a delicate scrape.
Note that the phenotype of the cells may change if they are cultured in 2D after their removal from the matrix.
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Yes, these products were designed to work together.
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Gibco Geltrex Matrix is a soluble form of reduced growth factor (RGF) basement membrane extract (BME) purified from murine Engelbreth-Holm-Swarm tumor. Historically these preparations have contained the lactate dehydrogenase-elevating virus (LDEV), which does not impact the performance of the material but may be of concern in certain applications such as xenografts or other clinically focused research. New manufacturing methods are now available that allow for the removal of this virus. The resulting LDEV-free material is tested using PCR methods to confirm that the material is virus-free.
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Gibco Geltrex Matrix and collagen (rat tail and bovine) can be used as either a coating solution or a 3D gel matrix. CELLstart Substrate was developed to be used as a xeno-free coating matrix for only ESC applications (as a substitute for Gibco Geltrex Matrix or Matrigel Matrix, which are of animal origin). AlgiMatrix Matrix is a 3D scaffold-type matrix that does not support cell attachment but does provide a good environment for growing spheroids that can be easily harvested for downstream applications.
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