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StemXVivo Serum-Free Human MSC Expansion Media

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Morphology of Human MSCs.
(2)
Phenotypic Analysis of Human MSCs Expanded in StemXVivo® Serum-Free Human Mesenchymal Stem Cell Expansion Media for 3 Passages.
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CCM014
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Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, MSCs are cultured in serum-free expansion media using the following protocol:

  • Plate MSCs on Fibronectin-coated culture dishes
  • Culture MSCs to 80-90% confluency in serum-free expansion media
  • Passage MSCs using fresh serum-free expansion media
 

 

Reagents Provided

Reagents supplied in the StemXVivo® Serum-Free Mesenchymal Stem Cell Expansion Media (Catalog # CCM014):

  • 250 mL of serum-free MSC expansion media

 

Other Supplies Required

Reagents

  • Penicillin-Streptomycin (100X)
  • Human Fibronectin (R&D Systems, Catalog # 1918-FN-02M)
  • Trypsin-EDTA (10X)
  • Phosphate-Buffered Saline (PBS)

Materials

  • Human MSCs
  • 75 cm2 tissue culture flasks
  • 15 mL centrifuge tubes
  • Pipettes and pipette tips
  • Serological pipettes

Equipment

  • 37 °C and 5% CO2 incubator
  • Centrifuge
  • Hemocytometer
  • Inverted microscope
  • 2 °C to 8 °C refrigerator
  • 37 °C water bath

 

Procedure Overview

Culturing Mesenchymal Stem Cells

Coat plates with 5 µg/mL Human Fibronectin.

Coat plates with 5 µg/mL Human Fibronectin

Add 4.5-5.0 x 105 MSCs resuspended in 20 mL of the pre-warmed StemXVivo® Serum-Free MSC Expansion Media to a T75 flask.

Detach the cells from the cell culture dish.

Replace the medium every 2-3 days with fresh StemXVivo® Serum-Free MSC Expansion Media.

Culture cells to 80-90% confluency.

Replace the medium every 2-3 days with fresh StemXVivo<sup>®</sup> Serum-Free MSC Expansion Media

Subculturing Mesenchymal Stem Cells

Remove and discard the media from the T75 tissue culture flask(s) containing the MSCs.

Wash the cells with PBS.

Transfer the cells to a cryovial.
 

Thawing of Cryopreserved Stem Cells

Add 3 mL of pre-warmed trypsin to the MSCs and incubate the flask at 37 °C.

Add 3 mL of pre-warmed trypsin to the MSCs and incubate the flask at 37 <sup>°</sup>C

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

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

Resuspend the cell pellet in 5 mL of StemXVivo® Serum-Free MSC Expansion Media.

Resuspend the cell pellet in 5 mL of StemXVivo<sup>®</sup> Serum-Free MSC Expansion Media

Perform a cell count..

Perform a cell count.

Add 4.5-5.0 x 105 MSCs resuspended in 20 mL of pre-warmed StemXVivo® Serum-Free MSC Expansion Media into each Fibronectin-coated T75 flask.

Culture the MSCs to 80-90% confluency.

Add 4.5-5.0 x 10<sup>5</sup> MSCs resuspended in 20 mL of pre-warmed StemXVivo<sup>®</sup> Serum-Free MSC Expansion Media into each Fibronectin-coated T75 flask

Citations for StemXVivo Serum-Free Human MSC Expansion Media (3)

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Product Documents for StemXVivo Serum-Free Human MSC Expansion Media

Certificate of Analysis

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