Human Embryonic Stem Cell Culture: The Next Step

Initial culturing techniques for human embryonic stem (hES) cells originated from methodology developed for murine ES cells, including the use of mouse embryonic fibroblast (MEF) feeder layers. While MEF feeder layers have proven to be a robust surface for long-term culture of hES cells, they present a number of limitations. Concerns about contamination of hES cells from animal-derived pathogens from MEF feeder layers makes this method of culturing unsuitable for clinical applications.

Recent studies have clearly delineated that hES cells can be grown in the absence of feeders as long as they are plated on an appropriate extracellular matrix (ECM) and cultured in feeder-conditioned media or media supplemented with appropriate soluble factors. In 2001, the first feeder-free hES culture system was demonstrated.

BD Matrigel Matrix coupled with different feeder-conditioned media has been widely accepted as an alternative method for culturing hES cells. Many ­independent studies suggest that the BD Matrigel Matrix surface will likely be compatible with a number of hES culturing media and that this surface may also be used to culture other types of ES cells.

When cultured on BD Matrigel Matrix, hES cells maintain normal karyotype and demonstrate a stable proliferation rate and high telomerase activity. These hES cells express characteristic undifferentiated hES cell markers, they form teratomas in severe combined immunodeficient (SCID) mice and differentiate into cells from all three germ layers.

Advantages of using BD Matrigel ­Matrix 6-well plates for human ES cell culture:

• Ready to use convenience
• Quality assurance tested product
• Optimized culture protocol
• Easier DNA, RNA & protein isolation
• Improved transfection efficiencies
• Maintenance of only one cell type
• Compatible with multiple media and ES cell lines