Cultrex ReadyBME is a pre-diluted and ready-to-use coating substrate that improves the consistency and efficiency of feeder-free pluripotent stem cell cultures.
What is Cultrex ReadyBME?
Cultrex ReadyBME is a prediluted and ready-to-use version (working concentration of 80-120 μg/mL) of Cultrex Reduced Growth Factor (RGF) BME, Stem Cell Qualified that is useful for promoting the attachment, expansion, and differentiation of human embryonic stem cells and induced pluripotent stem cell under feeder-free conditions. Pluripotent stem cells cultured on Cultrex ReadyBME maintain robust expression of the stemness markers, SSEA-4 and Oct-3/4. They also retain pluripotency as demonstrated by germ layer differentiation into ectoderm, mesoderm, and endoderm.
Cultrex Basement Membrane Extract (BME) is a soluble form of basement membrane purified from Engelbreth-Holm-Swarm (EHS) tumor. Cultrex BME gels at 37 °C to form a reconstituted basement membrane. The major components of BME include laminin, collagen IV, entactin, and heparin sulfate proteoglycan. These proteins derived from the extracellular matrix are frequently used to support the attachment, maintenance and expansion of stem cells, including human embryonic (ESCs) and induced pluripotent stem cells (iPSC) culture under feeder free conditions.
Cultrex ReadyBME Reduces Handling and Performance Variability
As a ready-to-use coating solution, Cultrex ReadyBME eliminates the need for careful thawing and handling required when using standard Cultrex BME formulations. It increases workflow efficiency and user-to-user consistency by being provided in a form that can be taken directly from the refrigerator and immediately used to coat cell culture surfaces.
Murine Engelbreth-Holm-Swarm (EHS) tumor
Tested following USP <71> sterility guidelines.
Cell Culture Testing
Promotes the attachment of human induced pluripotent stem cells (iPSCs).
Effectively maintains human iPSCs in a pluripotent state in a feeder-free culture.
Tested negative by PCR test for a total of 31 organisms and viruses, including: mycoplasma, 17 bacterial and virus strains typically included in mouse antibody production (MAP) testing, and 13 additional murine infectious agents including LDEV.
Maintenance of Human Induced Pluripotent Stem Cells.
JOY6 human induced pluripotent stem cell (hiPSC) colonies were successfully cultured for > 10 passages using Cultrex ReadyBME. A) Representative image of JOY6 hiPSC colonies cultured on Cultrex ReadyBME. B) hiPSCs cultured on Cultrex ReadyBME retain stemness markers as shown by flow cytometry using a Human/Mouse Oct-3/4 APC-conjugated Antibody (Catalog # IC1759A) and Human/Mouse SSEA-4 Fluorescein-conjugated Antibody (Catalog # FAB1435F). C) hiPSCs remain negative for SSEA-1expression as detected using a Human/Mouse SSEA-1 PerCP-conjugated Antibody (Catalog # FAB2155C).
Colony Formation of Human iPSC and ESC Cultured using Cultrex ReadyBME.
Three human induced pluripotent stem cell lines (IPSK3, IBJ6, and JOY6) and one human embryonic stem cell line (BG01V) were cultured using Cultrex ReadyBME. Representative images of colonies for each of the pluripotent stem cell lines shows normal colony formation. BG01V human embryonic stem cells are licensed for ViaCyte, Inc.
Differentiation of Human Induced Pluripotent Stem Cells.
IBJ6 human induced pluripotent stem cells were cultured for 6 passages using Cultrex ReadyBME. Pluripotency was tested using the Human Pluripotent Stem Cell Functional Identification Kit (Catalog # SC027B), which differentiates hiPSCs into the three germ layers.Immunocytochemistry showed that IBJ6 hiPSC successfully differentiated into endoderm (SOX17), mesoderm (Brachyury),and ectoderm (Otx2). Cells were counterstained with DAPI (Catalog # 5748).
Cultrex ReadyBME Performs Equal to Cultrex Stem Cell Qualified RGF BME.
Human induced pluripotent stem cells (iPSCs) were maintained in culture using Cultrex ReadyBME or Stem Cell Qualified (SCQ) Cultrex RGF BME (Catalog # 3434-005-02) and evaluated for (A) expression of stemness markers, SSEA-4 and Oct-3/4, and (B) differentiation capacity into endoderm. A) iPSCs cultured using Cultrex ReadyBME showed similar expression of SSEA-4 and Oct-3/4 compared to SCQ Cultrex RGF BME. B) iPSC cultured using Cultrex ReadyBME showed similar efficiency to differentiate into endoderm, as determined by SOX17 expression, compared to cells grown using SCQ Cultrex RGF BME
Preparation & Storage
Product is stable for 12 months from date of manufacture. See lot specific Certificate of Analysis for expiration date.
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended on the product label.
Cultrex-ReadyBME has worked very well for our lab. H9 hESCs (WA09) were grown on Cultrex-ReadyBME and was directly compared side-by-side to our normal basement membrane. So far, I’ve grown the cells for more the 5 passages using Accutase and haven’t observed any changes in proliferation rate, viability, cell morphology or the increased appearance of differentiated cells.
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