Related Experiment Video
Updated: Sep 20, 2026
Preparation of Mouse Embryonic Fibroblast Cells Suitable for Culturing Human Embryonic and Induced Pluripotent Stem Cells
Published on: June 21, 2012
Comparative evaluation of various human feeders for prolonged undifferentiated growth of human embryonic stem cells
Mark Richards1, Shawna Tan, Chui-Yee Fong
1Department of Obstetrics and Gynecology, National University of Singapore, Kent Ridge, Singapore.
Abstract:
Human embryonic stem (hES) cells are typically derived and serially propagated on inactivated murine embryonic fibroblast (MEF) feeders. The use of MEFs and other components of animal origin in the culture media for hES cell support substantially elevates the risk of contaminating these cell lines with infectious agents of animal origin thereby severely limiting their potential for clinical application. We have previously shown that it is possible to derive and establish new hES cell lines in a xeno-free culture system using human fetal muscle fibroblast feeders. In this report, we have comparatively evaluated a panel of 11 different human adult, fetal, and neonatal feeders for hES cell support and have ranked them as supportive and non-supportive. We report that two adult skin fibroblast cell lines established in-house from abdominal skin biopsies supported prolonged undifferentiated hES cell growth for over 30 weekly passages in culture. Furthermore, hES cell lines cultured on adult skin fibroblast feeders retain hES cell morphology and remain pluripotent. Also, differences in feeder support exist between human cell types and sources. The use of human adult skin feeders is convenient for hES cell support given the ease of obtaining skin biopsies.
Insights
Human embryonic stem cells (hESCs) can be cultured without animal products using human adult skin fibroblasts as feeders. This xeno-free method supports prolonged undifferentiated growth, enhancing safety for clinical applications.
Area of Science:
- Stem Cell Biology
- Regenerative Medicine
- Cell Culture Technology
Background:
- Human embryonic stem cells (hESCs) are typically cultured on animal-derived murine embryonic fibroblasts (MEFs).
- The use of animal-derived components poses a risk of contamination with infectious agents, limiting clinical applications.
- Previous research demonstrated xeno-free hESC derivation using human fetal muscle feeders.
Purpose of the Study:
- To comparatively evaluate human adult, fetal, and neonatal feeders for supporting hESC culture.
- To identify optimal human feeder cell types for xeno-free hESC propagation.
- To assess the suitability of adult skin fibroblasts for long-term, undifferentiated hESC growth.
Main Methods:
- Culturing hESCs on a panel of 11 different human feeder cell lines (adult, fetal, neonatal).
- Assessing feeder support based on hESC growth, morphology, and pluripotency.
- Ranking feeders as supportive or non-supportive for hESC culture.
- Serial propagation of hESCs for over 30 passages on selected feeders.
Main Results:
- Identified specific human adult skin fibroblast lines that effectively support prolonged undifferentiated hESC growth.
- Demonstrated that hESCs cultured on adult skin feeders maintain their characteristic morphology and pluripotency.
- Showcased significant differences in feeder support capabilities among various human cell types and sources.
- Highlighted the convenience and efficacy of using adult skin fibroblasts derived from easily obtainable biopsies.
Conclusions:
- Human adult skin fibroblasts provide a robust and xeno-free feeder system for hESC culture.
- This approach mitigates risks associated with animal-derived materials, paving the way for safer clinical translation.
- Adult skin fibroblasts represent a practical and effective alternative to MEFs for maintaining hESC pluripotency and proliferation.
