Proliferative feeder cells support prolonged expansion of human embryonic stem cells

Area of Science:

• Stem Cell Biology
• Developmental Biology

Background:

• Long-term human embryonic stem (hES) cell culture typically relies on mitotically inactive mouse embryonic fibroblasts (mEFs) as feeder layers.
• Maintaining hES cell pluripotency during prolonged expansion is crucial for research and therapeutic applications.

Purpose of the Study:

• To investigate the potential of using proliferative mEFs as feeder cells for sustained hES cell expansion.
• To determine if proliferative mEFs can maintain the undifferentiated state of hES cells during long-term culture.

Main Methods:

• Undifferentiated hES cell colonies were cultured on six different densities of proliferative mEFs.
• Cell cultures were maintained for over 130 days.
• Pluripotency and differentiation status were assessed.

Main Results:

• hES cell colonies cultured on proliferative mEFs at densities of 1 x 10(5) to 5 x 10(5) cells successfully proliferated.
• These colonies remained undifferentiated throughout the 130-day culture period.
• The proliferative mEF feeder layer supported hES cell propagation similarly to mitotically inactive mEFs.

Conclusions:

• Specific densities of proliferative mEFs can effectively support the long-term propagation and self-renewal of undifferentiated hES cells.
• This method offers a potential alternative for hES cell culture and may aid in identifying essential cytokines and adhesion molecules for pluripotency maintenance.