StemPro hESC SFM from Life Technologies

April 24, 2013

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McLean Hospital/Harvard Medical School
Molecular Neurobiology Lab
Post-doc

hES (H9) cells cultivated under feeder-free conditions with StemPro media.

Company:

Life Technologies

Product Name:

StemPro Media

Catalog Number:

A1000701
Image

StemPro media is a fully-defined, serum- and feeder-free medium (SFM) specially formulated for the growth and expansion of human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). Since Dr. Yamanaka developed the breakthrough technology of generating iPSC by using 4 factors, a lot of research has focused on its application. One of the biggest challenges with this research was contamination of feeder cells from mouse and other undefined factors from serum during cell expansion and maintenance. In addition, in order to perform functional assays with hES/hiPSC, feeder cells must be eliminated. For these reasons, a fully-defined, serum- and feeder-free medium is necessary for hES/hiPSC experiments.

Experimental Design and Results Summary

Application

Growth and expansion of hESCs and hiPSCs without feeder cells.

Starting Material

Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs).

Protocol Overview

1. coat culture dishes with Geltrex at least 1 hr before cell seeding. 2. Gently add pre-warmed collagenase IV or dispase to cells and incubate 5 min. 3. Dissociate cells with StemPro media by pipetting; pellet cells by centrifuging. 4. Aspirate media and resuspend cell pellet with 1-2mL StemPro media by pipetting. 5. Gently seed on the Geltrex coated dishes. 6. Incubate until cells are attached on the plate. 7. Add more media.

Tips

For feeder-free culture conditions, culture dishes should be coated with Geltrex (Life Technoloies, #A1413301).

Results Summary

After several passages under feeder-free conditions, the pluripotency of hESC/hiPSCs were checked by alkaline phosphate staining (AP) and immunostaining with anti-Oct4, Sox2, SSEA-4, and Tra1-60 antibodies. Staining was positive for all 4 markers, which implied pluripotency of hESC/hiPSCs after cultivation under feeder-free culture.

Additional Notes

bFGF which is not included in the media is still necessary for hES/hiPSC culture.

Image Gallery

hES (H9) cells cultivated under feeder-free conditions with StemPro media.

Summary

The Good

Good media for hES/hiPSC culture under feeder-free conditions.

The Bad

A little expensive compared to traditional hES/hiPSC feeder-free culture media.

The Bottom Line

Fully-defined, serum- and feeder-free medium is required for clinical applications of hES/hiPSC technology. StemPro media is a good option for moving hES/hiPSC into the clinic.

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