Establishment and characterization of immortalized mouse embryonic fibroblasts by SV40 large T antigen
- 1National Collection of Authenticated Cell Cultures, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai, China
- 2University of Chinese Academy of Sciences, Beijing, China
Abstract
Mouse embryonic fibroblasts (MEFs), which are typically derived from mouse embryos between embryonic days 12.5 and 13.5, serve as a widely used feeder layer for pluripotent stem cell culture. MEFs exhibit a limited proliferative capacity and are typically used within five passages after which they undergo early senescence. However, the repeated preparation of primary MEFs introduces batch-to-batch variability and raises ethical concerns regarding animal use, thereby limiting the reproducibility and scalability of stem cell culture. To overcome these limitations, here we established a nontumorigenic immortalized MEF line (designated as SV40-CMEF) from C57BL/6J mice via SV40 large T antigen (SV40LT) transfection. SV40-CMEFs showed genomic integration, enhanced proliferation, and the capacity to support the self-renewal of R1/E mouse embryonic stem cells (mESC) as feeder cells. They also retained their tri-lineage differentiation potential. Notably, SV40-CMEFs did not form colonies in soft agar or tumors in immunodeficient mice. Transcriptomic profiling revealed substantial global remodeling compared with that in primary MEFs; however, functional gene modules associated with feeder capacity, fibroblast identity, and proliferation were selectively retained, whereas senescence markers were suppressed. This cell line provides a convenient, well-characterized resource for stem cell culture and cell differentiation research.
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BibTeX
@article{li2026establishment,
title = {{Establishment and characterization of immortalized mouse embryonic fibroblasts by SV40 large T antigen}},
author = {Xin Li and Ting Shao and Lan Xu and Li Zheng and Yue-Lei Chen},
journal = {Frontiers in Cell and Developmental Biology},
year = {2026},
month = oct,
doi = {10.3389/fcell.2026.1936956},
url = {https://doi.org/10.3389/fcell.2026.1936956},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Establishment and characterization of immortalized mouse embryonic fibroblasts by SV40 large T antigen
AU - Xin Li
AU - Ting Shao
AU - Lan Xu
AU - Li Zheng
AU - Yue-Lei Chen
PY - 2026
DA - 2026/10/22
JO - Frontiers in Cell and Developmental Biology
DO - 10.3389/fcell.2026.1936956
UR - https://doi.org/10.3389/fcell.2026.1936956
AB - Mouse embryonic fibroblasts (MEFs), which are typically derived from mouse embryos between embryonic days 12.5 and 13.5, serve as a widely used feeder layer for pluripotent stem cell culture. MEFs exhibit a limited proliferative capacity and are typically used within five passages after which they undergo early senescence. However, the repeated preparation of primary MEFs introduces batch-to-batch variability and raises ethical concerns regarding animal use, thereby limiting the reproducibility and scalability of stem cell culture. To overcome these limitations, here we established a nontumorigenic immortalized MEF line (designated as SV40-CMEF) from C57BL/6J mice via SV40 large T antigen (SV40LT) transfection. SV40-CMEFs showed genomic integration, enhanced proliferation, and the capacity to support the self-renewal of R1/E mouse embryonic stem cells (mESC) as feeder cells. They also retained their tri-lineage differentiation potential. Notably, SV40-CMEFs did not form colonies in soft agar or tumors in immunodeficient mice. Transcriptomic profiling revealed substantial global remodeling compared with that in primary MEFs; however, functional gene modules associated with feeder capacity, fibroblast identity, and proliferation were selectively retained, whereas senescence markers were suppressed. This cell line provides a convenient, well-characterized resource for stem cell culture and cell differentiation research.
SN - 2296-634X
ER -