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StemXVivo Culture Matrix (100X)

Catalog #: CCM013
2 Citations Datasheet / COA / SDS

Discontinued Product

CCM013 has been discontinued.
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StemXVivo Culture Matrix (100X) Summary

Kit Summary

A defined matrix for the growth and differentiation of stem cells and progenitor cells.

Key Benefits

  • Reduces experimental variation versus undefined alternatives
  • Higher lot-to-lot consistency versus cellular extract preparations
  • Quick and simple coating procedure

CCM013 Data Examples
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Why is it important to use a defined culture matrix?

A critical concern when using long-term cell culture protocols is minimizing the amount of introduced experimental variability.

Many researchers plate a variety of cell types using solubilized basement membrane extracts (BME) from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma. These preparations are known to include an undetermined number of extracellular matrix proteins and growth factors. To provide a more defined alternative, R&D Systems optimized a proprietary mixture of recombinant human adhesion molecules specifically for the culture of stem cells and progenitor cells.

StemXVivo® Culture Matrix:

  • Reduces experimental variation versus undefined alternatives.
  • Has greater lot-to-lot consistency versus cellular extract preparations.
  • Is specifically designed for stem cell research.
  • Supports growth of TE03, CyT49s, and BG01V human embryonic stem cells, iPS2 human induced pluripotent stem cells, rat cortical stem cells, and mesenchymal stem cells.
  • Requires a quick and simple coating procedure.
  • Provides a defined alternative to EHS-BME in feeder-free stem cell culture.
 

 

Matrix Components

StemXVivo® Culture Matrix is a defined proprietary mixture of R&D Systems® premium quality recombinant human adhesion molecules.

Provided as 1 mL (100X). Dilute 1:100 prior to use. For a 6-well tissue culture plate, 1.5 mL is needed to coat each well. For a 24-well tissue culture plate, or to coat coverslips in a 24-well tissue culture plate, 400 µL is needed per well.

StemXVivo® Culture Matrix is mycoplasma negative as tested by the MycoProbe™ Mycoplasma Detection Kit (Catalog # CUL001B) and has low Endotoxin levels (1X solution < 1 EU/mL).

 

Stability and Storage

Upon receipt, the StemXVivo® Culture Matrix should be stored at < 20 °C in a manual defrost freezer. The culture matrix should be thawed at 2 °C – 8 °C before use. Thawed culture matrix can be stored at 2°C – 8 °C for up to 1 month or aliquoted and stored at < 20 °C for up to 3 months.

 

Data Examples
StemXVivo Culture Matrix Supports the Growth of TE03 Human Embryonic Stem Cells.
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StemXVivo® Culture Matrix Supports the Growth of TE03 Human Embryonic Stem Cells.TE03 human embryonic stem cells were grown for 10 days on StemXVivo® Culture Matrix (Catalog # CCM013) or basement membrane extracts (BME) from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma. The cells were fixed and stained after passage 1 (5 days for passage 0 and 5 days for passage 1). Selective p160 ROCK inhibitor Y27632 (10 µM, Catalog # 1254) was added to the cultures for the first 24 hours of each passage. The cells were double stained using an Anti-TRA-1-81 antibody (green) to detect undifferentiated cells and an Anti-Laminin antibody (red) to detect differentiated cells. Nuclei were counterstained with DAPI (blue). Similar marker expression and degree of spontaneous differentiation was detected in cells grown on the two matrices. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
StemXVivo Culture Matrix Supports the Growth of TE03 Human Embryonic Stem Cells.
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StemXVivo® Culture Matrix Supports the Growth of TE03 Human Embryonic Stem Cells. TE03 human embryonic stem cells were grown for 10 days on StemXVivo® Culture Matrix (Catalog # CCM013) or basement membrane extracts (BME) from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma. The cells were fixed and stained after passage 1 (5 days for passage 0 and 5 days for passage 1). Selective p160 ROCK inhibitor Y27632 (10 mM, Catalog # 1254) was added to the cultures for the first 24 hours of each passage. The cells were double stained using a Rat Anti-Human/Mouse Oct-3/4 Monoclonal Antibody (MAB1759; red) to detect undifferentiated cells and an Anti-Nestin Polyclonal Antibody (green) to detect differentiated cells. Nuclei were counterstained with DAPI (blue). Similar marker expression and degree of spontaneous differentiation was detected in cells grown on the two matrices. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Data Courtesy of Dr. Ron McKay, Lieber Institute
StemXVivo Culture Matrix Supports the Growth of CyT49s Human Embryonic Stem Cells.
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StemXVivo® Culture Matrix Supports the Growth of CyT49s Human Embryonic Stem Cells. CyT49s human embryonic stem cells were grown on StemXVivo® Culture Matrix (Catalog # CCM013) or basement membrane extracts (BME) from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma. Cell counts at passage 0 (p0), p1, p2, and p3 indicated a similar growth rate for cells cultured on the two matrices. In addition, to its recommended working dilution of 1:100, StemXVivo® Culture Matrix (Catalog # CCM013) was applied at 1:200, with comparable results.
iPS2: Nectin (Red), DAPI (Blue)
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StemXVivo® Culture Matrix Supports the Growth and Differentiation of BG01V Human Embryonic Stem Cells. BG01V human embryonic stem cells were grown for 3 passages on StemXVivo® Culture Matrix (Catalog # CCM013) then randomly differentiated by growth factor withdrawal for 10 days. To confirm that growth on this matrix supported pluripotent differentiation, cells were stained with a Goat Anti-Rat Nestin Antigen Affinity-purified Antibody (Catalog # AF2736, ectoderm marker), a Mouse Anti-Human alpha-Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420, mesoderm marker), and a Goat Anti-Human SOX17 Antigen Affinity-purified Antibody (Catalog # AF1924, endoderm marker). The cells were stained using the NorthernLights™ 557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001; red) or NorthernLights 557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL007; red) and the nuclei were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
StemXVivo Culture Matrix Supports the Growth iPS2 Human Induced Pluripotent Stem Cells
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StemXVivo® Culture Matrix Supports the Growth iPS2 Human Induced Pluripotent Stem Cells. iPS2 human induced pluripotent stem cells were grown on StemXVivo® Culture Matrix (Catalog # CCM013). To confirm the expected expression of pluripotency markers, SSEA-4 and Oct-3/4 were detected using a Human/Mouse SSEA-4 Monoclonal Antibody (Catalog # MAB1435) and a Human Oct-3/4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1759). Cells were stained using the NorthernLights™ 493-conjugated Donkey Anti-Mouse IgG Secondary Antibody (green; Catalog # NL009) and the NorthernLights™ 557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001; red) and nuclei were counterstained with DAPI (blue). High expression of SSEA-4 and Oct-3/4 supports the pluripotent status of this cell population. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
StemXVivo Culture Matrix Supports the Growth iPS2 Human Induced Pluripotent Stem Cells
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StemXVivo® Culture Matrix Supports the Growth iPS2 Human Induced Pluripotent Stem Cells. iPS2 human induced pluripotent stem cells were grown on StemXVivo® Culture Matrix (Catalog # CCM013). To confirm the expected expression of cell markers, Oct-4A and E-Cadherin were detected using a Mouse Anti-Human Oct-4A Monoclonal Antibody (Catalog # MAB17591) and a Goat Anti-Human E-Cadherin Affinity-purified Polyclonal Antibody (Catalog # AF648). The cells were stained using the NorthernLights™ 493-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL009; green) and the NorthernLights™ 557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001; red) and nuclei were counterstained with DAPI (blue). High expression of the Oct-4A isoform and E-Cadherin supports the pluripotent status of this cell population. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Specifications

Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Species
Human

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Assay Procedure

Refer to the product datasheet for complete assay procedure.

 

 

  1.   Thaw StemXVivo Culture Matrix at 2 °C to 8 °C before use.
  2.   Determine the volume of diluted culture matrix required. For example, 1.5 mL is needed for each well of a 6-well tissue culture plate. For a 24-well tissue culture plate, or to coat coverslips in a 24-well tissue culture plate, 400 mL is needed per well.
  3.   Dilute the Culture Matrix 1:100 in sterile 1X PBS. Mix gently. Do not vortex.

    Note:The concentration of culture matrix used may need to be optimized for some cells
  4.   Immediately add diluted culture matrix to the plates or coverslips.
  5.   Incubate at 37 °C for 2 - 3 hours.
  6.   Immediately prior to plating the cells, remove the diluted culture matrix, and rinse once with sterile 1X PBS.
  7.   Plate the cells.
  8.   Passaging of the cells can be achieved using standard methods; either enzymatic or manual.

 

 

Thaw
Dilute
Add to Plates or Coverslips
Remove Immediately Before Plating Cells
Passaging

Citations for StemXVivo Culture Matrix (100X)

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Endocrine fibroblast growth factor FGF19 promotes prostate cancer progression.
    Authors: Feng S, Dakhova O, Creighton C, Ittmann M
    Cancer Res, 2013-02-25;73(8):2551-62.  2013-02-25
  2. Wnt1 and BMP2: two factors recruiting multipotent neural crest progenitors isolated from adult bone marrow.
    Authors: Glejzer A, Laudet E, Leprince P, Hennuy B, Poulet C, Shakhova O, Sommer L, Rogister B, Wislet-Gendebien S
    Cell. Mol. Life Sci., 2010-10-26;68(12):2101-14.  2010-10-26

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