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Updated: Nov 6, 2025
Collection of Serum- and Feeder-free Mouse Embryonic Stem Cell-conditioned Medium for a Cell-free Approach
Published on: January 8, 2017
Feeder-Dependent/Independent Mouse Embryonic Stem Cell Culture Protocol
Hatice Burcu Şişli1, Selinay Şenkal1, Derya Sağraç1
1Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, Istanbul, Turkey.
Abstract:
Mouse embryonic stem cells (mESCs) were first derived and cultured nearly 30 years ago and have been beneficial tools to create transgenic mice and to study early mammalian development so far. Fibroblast feeder cell layers are often used at some stage in the culture protocol of mESCs. The feeder layer-often mouse embryonic fibroblasts (MEFs)-contribute to the mESC culture as a substrate to increase culture efficiency, maintain pluripotency, and facilitate survival and growth of the stem cells. Various feeder-dependent and feeder-independent culture and differentiation protocols have been established for mESCs. Here we describe the isolation, culture, and preparation feeder cell layers and establishment of feeder-dependent/independent protocol for mESC culture. In addition, basic mESC protocols for culture, storage, and differentiation were described.
Insights
Mouse embryonic stem cells (mESCs) require fibroblast feeder cells for optimal culture. This study details protocols for feeder cell preparation and establishes feeder-dependent and independent methods for mESC culture, storage, and differentiation.
Area of Science:
- Stem Cell Biology
- Developmental Biology
- Mammalian Genetics
Background:
- Mouse embryonic stem cells (mESCs) are crucial for studying early mammalian development and creating transgenic models.
- Fibroblast feeder layers, typically mouse embryonic fibroblasts (MEFs), are commonly used to support mESC culture by enhancing efficiency, pluripotency, survival, and growth.
Purpose of the Study:
- To describe the isolation, culture, and preparation of feeder cell layers for mESC culture.
- To establish feeder-dependent and feeder-independent protocols for mESC culture.
- To outline basic mESC protocols for culture, storage, and differentiation.
Main Methods:
- Isolation and culture of mouse embryonic fibroblasts (MEFs) as feeder layers.
- Establishment of feeder-dependent mESC culture systems.
- Development of feeder-independent mESC culture protocols.
- Standardization of mESC culture, storage, and differentiation procedures.
Main Results:
- Successful isolation and preparation of functional feeder cell layers.
- Demonstration of effective feeder-dependent and feeder-independent mESC culture protocols.
- Established protocols for mESC storage and differentiation.
Conclusions:
- Feeder cell layers play a vital role in maintaining mESC pluripotency and viability.
- Both feeder-dependent and feeder-independent methods can be successfully employed for mESC culture.
- The described protocols provide a comprehensive resource for mESC manipulation and research.
