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A hollow sphere soft lithography approach for long-term hanging drop methods
Won Gu Lee1, Daniel Ortmann, Matthew J Hancock
1Department of Medicine, Center for Biomedical Engineering, Harvard Medical School, Brigham and Women's Hospital, Boston, Massachusetts, USA.
Tissue Engineering. Part C, Methods
|June 10, 2009
Summary
A novel hollow sphere method enables long-term embryonic stem cell culture in hanging drops (HD). This technique supports larger embryoid body growth and differentiation for regenerative medicine applications.
Area of Science:
- Stem Cell Biology
- Biomaterials Engineering
- Regenerative Medicine
Background:
- Conventional hanging drop (HD) methods for embryonic stem cell (ESC) aggregates or embryoid bodies (EBs) are limited by small droplet volumes (<50 microL).
- These limitations restrict culture duration to a few days due to rapid media depletion and inconsistent conditions, hindering long-term studies and applications.
Purpose of the Study:
- To develop a modified HD method enabling long-term (10-15 days) ESC culture with enhanced media conditions.
- To investigate the efficacy of hollow sphere (HS) structures as improved cell culture chambers for EB growth.
Main Methods:
- Fabrication of hollow sphere (HS) structures using noncured poly(dimethylsiloxane) (PDMS) mixtures.
- Culturing embryoid bodies (EBs) within HS structures providing a larger media reservoir (500 microL) in a HD format.
Main Results:
- EBs cultured in HS structures demonstrated approximately double the size compared to conventional HD methods after 10 days.
- HS cultures maintained consistent media conditions, preventing depletion over the extended culture period.
- Multilineage differentiation capacity of EBs was preserved in HS cultures, comparable to conventional HD methods.
Conclusions:
- The developed HS-based HD method facilitates long-term embryonic stem cell culture with improved growth and differentiation.
- This approach offers a robust platform for stem cell culture, potentially benefiting regenerative medicine strategies.
- The ease of fabrication and enhanced culture features make HS structures a promising tool for stem cell research.
