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StemXVivo Xeno-Free Human MSC Expansion Media

R&D Systems, part of Bio-Techne | Catalog # CCM021

R&D Systems, part of Bio-Techne

Key Product Details

Morphology of Mesenchymal Stem Cells (MSCs) Cultured in StemXVivo Xeno-Free Human MSC Expansion Media.
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Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, mesenchymal stem cells can be analyzed for marker expression using the following procedure:

  • Resuspend cells in pre-warmed StemXVivo® Xeno-Free Human MSC Expansion Media
  • Plate the cells on Fibronectin-coated plates
  • Culture the cells at 37 °C and 5% CO2
 

 

Reagents Provided

Reagents supplied in the StemXVivo® Xeno-Free Human MSC Expansion Media (Catalog # CCM021)::

  • 500 mL of Xeno-Free Human MSC expansion media

 

Other Supplies Required

Reagents and Materials

  • Bone marrow-derived MSCs
  • Recombinant Human Fibronectin, ACFP (Catalog # ACFP4305)
  • Penicillin-Streptomycin (100X)
  • TrypLE™ Express (Invitrogen® or equivalent)
  • Phosphate-Buffered Saline (PBS)
  • 75 cm2 tissue culture flasks
  • 15 mL centrifuge tubes
  • Serological pipettes
  • Pipettes and pipette tips

Equipment

  • 37 °C and 5% CO2 humidified incubator
  • Centrifuge
  • Hemocytometer
  • Inverted microscope
  • 37 °C water bath

 

Protocol Overview

R&D Systems Protocol for Expanding and Subculturing MSCs in StemXVivo® Xeno-Free MSC Expansion Media (Catalog # CCM021)

Culturing Mesenchymal Stem Cells

Coat plates with Recombinant Human Fibronectin.

Coat plates with Recombinant Human Fibronectin.

Pre-warm the required amount of StemXVivo® Xeno-Free Human MSC Expansion Media to room temperature.

Pre-warm the required amount of media.

Resuspend 4.5-5.0 x 105 cells in 20 mL of the pre-warmed media.

Add the cell suspension to a Fibronectin-coated T75 flask.

Incubate the cells at 37 °C and 5% CO2 in a humidified atmosphere.

Replace spent media every 2-3 days with 20 mL of fresh, pre-warmed StemXVivo® Xeno-Free Human MSC Expansion Media.

Subculture the cells when they reach 80-90% confluency. Do not let the cultures become 100% confluent.

Resuspend cells in 20 mL of the pre-warmed media.
 

Subculturing Mesenchymal Stem Cells

Coat the plates with Recombinant Human Fibronectin.

Coat the plates with Recombinant Human Fibronectin.

Pre-warm StemXVivo® Xeno-Free Human MSC Expansion Media and TrypLE Express™ to room temperature.

Remove and discard the media from the flask(s).

Wash the cells once with PBS.

Detach the cells using TrypLE™ Express.

Pre-warm media.

Add 5 mL of StemXVivo® Xeno-Free Human MSC Expansion Media to the flask.

Transfer the cells to a 15 mL conical tube and centrifuge at 400 x g for 5 minutes.

Aspirate off the liquid.

Resuspend the cell pellet in a small amount of pre-warmed StemXVivo® Xeno-Free Human MSC Expansion Media.

Add 5 mL media to the flask.

Perform a cell count.

Perform a cell count.

Resuspend 4.5-5.0 x 105 cells in 20 mL of the pre-warmed StemXVivo® Xeno-Free Human MSC Expansion Media.

Add the cell suspension to a Fibronectin-coated T75 flask.

Incubate the cells at 37 °C and 5% CO2 in a humidified atmosphere.

Resuspend cells in pre-warmed media.
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