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Updated: Jan 10, 2026
Murine Dermal Fibroblast Isolation by FACS
Published on: January 7, 2016
Dermal fibroblast cultures recapitulate differences between deermice and mice in their responses to a Toll-like
Jonathan V Duong1, Aqsa Motiwala1, William J Hotz1
1Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine, Irvine, CA, United States.
Introduction:
The white-footed deermouse Peromyscus leucopus is a primary reservoir for the agents of Lyme disease and other zoonoses in North America and manifests infection tolerance for the bacteria, protozoa, and viruses it hosts. In previous in vivo studies, P. leucopus and Mus musculus differed in the degree of sickness and profiles of biomarkers after exposure to a bacterial lipopolysaccharide, a TLR4 agonist.
Methods:
As an approach for assessing immunity of mammals in nature and for longitudinal studies of colony animals in the laboratory, we evaluated primary dermal fibroblast cultures of P. leucopus and M. musculus in their short-term responses to a TLR2 agonist lipopeptide using bulk and single-cell RNA-seq.
Results:
By single-cell RNA-seq, cultures of both species comprised at least two types of fibroblasts, which were further differentiated in their responses to TLR agonists. With continued passage, the mouse cell population lost viability, while the deermouse cell population spontaneously transformed into a cell line stably maintained under standard conditions. Bulk RNA-seq revealed distinctive profiles for deermouse and mouse cells in arginine metabolism gene expression, high baseline transcription of the antioxidant transcription factor Nfe2l2 (Nrf2) in deermouse fibroblasts, and the transcription of the aging-associated cytokine interleukin-11 in agonist-treated mouse fibroblasts but not deermouse fibroblasts. In the cultures of both species, there was increased transcription of several types of endogenous retrovirus (ERV) and transposable elements (TEs) after exposure to the agonist. The transcribed ERV/TE sequences in M. musculus cells were generally longer in length and had greater potential for translation than sequences in treated P. leucopus cells.
Discussion:
The results indicate the feasibility of this in vitro model for both laboratory- and field-based studies, and that inherent differences between deer mice and mice in cell-autonomous innate immune responses and ERV/TE activation can be demonstrated in dermal fibroblasts as well as the animals themselves.
Insights
White-footed deermice fibroblasts offer a stable cell line for studying innate immunity and host-pathogen interactions, unlike mouse cells. This research reveals distinct immune responses and endogenous retrovirus activation in deermice versus mice.
Area of Science:
- Immunology
- Cell Biology
- Genomics
Background:
- White-footed deermice ( *Peromyscus leucopus* ) are key reservoirs for zoonotic diseases like Lyme disease.
- Deermice exhibit unique infection tolerance compared to laboratory mice (*Mus musculus*).
- Previous studies showed differential responses to lipopolysaccharide (LPS), a TLR4 agonist, between *P. leucopus* and *M. musculus*.
Purpose of the Study:
- To evaluate primary dermal fibroblast cultures from *P. leucopus* and *M. musculus* for assessing mammalian immunity in vitro.
- To compare short-term responses of deermouse and mouse fibroblasts to a TLR2 agonist using RNA sequencing.
Main Methods:
- Primary dermal fibroblast cultures from *P. leucopus* and *M. musculus* were established.
- Cells were stimulated with a TLR2 agonist (lipopeptide).
- Bulk and single-cell RNA sequencing (RNA-seq) were employed to analyze gene expression.
Main Results:
- Single-cell RNA-seq identified distinct fibroblast subtypes and differential responses to TLR agonists in both species.
- *P. leucopus* fibroblasts formed a stable cell line, whereas *M. musculus* cells lost viability with passage.
- Distinctive gene expression profiles were observed, including differences in arginine metabolism, Nrf2 transcription, and aging-associated cytokines.
- Both species showed increased transcription of endogenous retroviruses (ERVs) and transposable elements (TEs) upon agonist stimulation, with *M. musculus* exhibiting longer and more translatable sequences.
Conclusions:
- In vitro dermal fibroblast models are feasible for studying innate immunity and host-pathogen interactions in deermice and mice.
- Significant differences exist in cell-autonomous innate immune responses and ERV/TE activation between *P. leucopus* and *M. musculus* fibroblasts.
- These findings highlight inherent biological distinctions relevant to zoonotic disease research.
