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Cultrex ReadyBME, Stem Cell Qualified, Reduced Growth Factor

R&D Systems | Catalog # 3434-050-RTU

Ready-to-use Cultrex Basement Membrane Extract (BME) for the feeder-free culture of pluripotent stem cells.
R&D Systems
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Key Product Details

Features

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.

Cultrex ReadyBME, Stem Cell Qualified, Reduced Growth Factor (Catalog # 3434-050-RTU) is comparable to Geltrex™ hESC-Qualified, Ready-To-Use, Reduced Growth Factor Basement Membrane Matrix (Catalog # A1569601).

Key Benefits

  • Ready-to-use, stem cell qualified coating matrix
  • Minimizes handling and dilution variability
  • Improves experimental consistency and success
  • Reduces preparation time for feeder-free stem cell culture
  • Pre-diluted from concentrated Cultrex BME, Stem Cell Qualified
  • 2 EU/mL of Endotoxin

Species

Mouse

Product Summary for Cultrex ReadyBME, Stem Cell Qualified, Reduced Growth Factor

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.
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Product Specifications

Source

Murine Engelbreth-Holm-Swarm (EHS) tumor

Sterility Testing

No bacterial or fungal growth detected following 14 days in culture

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.

Viral Testing

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.

Scientific Data Images for Cultrex ReadyBME, Stem Cell Qualified, Reduced Growth Factor

Maintenance of Human Induced Pluripotent Stem Cells_Cultrex ReadyBME_3434-050-RTU

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).

Human iPSC and ESC Colony Formation_Cultrex ReadyBME_3434-050-RTU

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.
Trilineage Differentiation of Human Pluripotent Stem Cells using Cultrex ReadyBME

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 Similar Performance to Stem Cell Qualified Cultrex RGF BME

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

Formulation, Preparation, and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended on the product label.

Storage

Store the unopened product at 2 - 8 °C.

Stability

Product is stable for 12 months from date of manufacture. See lot specific Certificate of Analysis for expiration date.

Background: Basement Membrane Extracts

Basement membrane extract (BME) is a type of extracellular matrix (ECM) derived from Engelbreth-Holm-Swarm mouse sarcomas. BME is commonly used in cell culture and tissue engineering to provide a 2-D or 3-D scaffold for cells, allowing them to grow, differentiate, and interact in an environment that closely mimics the native ECM. This makes BME invaluable for modeling the behavior of cells in vivo.  All Cultrex BME formulations are rigorously tested to ensure performance, quality, and lot-to-lot consistency.

Additional Basement Membrane Extracts Products

Product Documents for Cultrex ReadyBME, Stem Cell Qualified, Reduced Growth Factor

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Citations for Cultrex ReadyBME, Stem Cell Qualified, Reduced Growth Factor

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  • Cultrex ReadyBME, Stem Cell Qualified, Reduced Growth Factor
    Name: Anonymous
    Verified Customer | Posted 09/15/2020
    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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