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Comparative Study of Reprogramming Efficiency and Regulatory Mechanisms of Placental- and Fibroblast-Derived Induced Pluripotent Stem Cells (iPSCs) in Mules.
Abstract: As an interspecies hybrid inheriting genetic material from horse and donkey lineages, mules provide a unique model for studying allele-specific regulatory dynamics. Here, we isolated adult fibroblasts (AFs) and placental fibroblasts (PFs) from mule tissues and reprogrammed them into induced pluripotent stem cells (iPSCs). Intriguingly, placental fibroblast-derived iPSCs (mpiPSCs) exhibited reduced reprogramming efficiency compared to adult fibroblast-derived iPSCs (maiPSCs). Through allele-specific expression (ASE) analysis, we systematically dissected transcriptional biases in parental cell types and their reprogrammed counterparts, revealing conserved preferential expression of asinine alleles in core pluripotency regulators (e.g., /, , ) across both cell lineages. Strikingly, mpiPSCs displayed stronger asinine allele dominance than maiPSCs, suggesting tissue-specific parental genomic imprinting. Mechanistic exploration implicated PI3K-AKT signaling as a potential pathway mediating the reprogramming inefficiency in placental fibroblasts. By integrating transcriptomic profiling with ASE technology, this study uncovers allele selection hierarchies during somatic cell reprogramming in hybrids and establishes a framework for understanding how parental genomic conflicts shape pluripotency establishment. These findings advance interspecies iPSC research by delineating allele-specific regulatory networks and providing insights into the molecular constraints of hybrid cellular reprogramming.
Publication Date: 2025-08-19 PubMed ID: 40864824PubMed Central: PMC12384536DOI: 10.3390/cimb47080671Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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Overview
- This study compares the efficiency and regulatory mechanisms of induced pluripotent stem cells (iPSCs) derived from placental fibroblasts and adult fibroblasts in mules, an interspecies hybrid of horses and donkeys.
- It investigates how genetic material from two species is selectively expressed and how this affects the reprogramming process to pluripotency, highlighting differences based on cell origin and allele-specific expression patterns.
Background and Model
- Mules are hybrids resulting from crossing a horse (Equus caballus) with a donkey (Equus asinus), inheriting genomes from both species.
- This interspecies hybrid status makes them an excellent model to study allele-specific regulatory dynamics, especially in contexts such as cellular reprogramming.
- Induced pluripotent stem cells (iPSCs) are somatic cells that have been genetically reprogrammed to a pluripotent state, capable of differentiating into various cell types.
- Reprogramming somatic cells into iPSCs helps investigate developmental biology, disease modeling, and therapeutic potentials.
Experimental Design
- Researchers isolated two types of fibroblasts from mules:
- Adult fibroblasts (AFs)
- Placental fibroblasts (PFs)
- Both cell types were reprogrammed into iPSCs, termed:
- maiPSCs – adult fibroblast-derived iPSCs
- mpiPSCs – placental fibroblast-derived iPSCs
- Their reprogramming efficiencies were compared to see how well placental and adult fibroblasts convert into iPSCs.
- Allele-specific expression (ASE) analysis was performed to identify preferential expression of horse or donkey alleles in parental and reprogrammed cells.
Key Findings
- Reprogramming Efficiency:
- Placental fibroblasts reprogrammed less efficiently than adult fibroblasts, indicating tissue-specific differences in cellular plasticity.
- Allele-Specific Expression and Parental Biases:
- Both adult and placental fibroblasts and their derived iPSCs showed a conserved preferential expression of asinine (donkey) alleles in key pluripotency regulator genes.
- Key regulatory genes involved include core pluripotency factors (though specific gene names were omitted in the abstract).
- Placental fibroblast-derived iPSCs exhibited stronger asinine allele dominance compared to adult-derived iPSCs, suggesting tissue-specific parental genomic imprinting influences gene expression during reprogramming.
- Mechanistic Insights:
- Pathway analysis indicated that the PI3K-AKT signaling pathway may be involved in the reduced reprogramming efficiency seen in placental fibroblasts.
- This implicates specific molecular signaling pathways as constraints during the reprogramming of certain cell types.
Scientific Significance
- The study delineates how parental genomes from two species interact and are differentially expressed in hybrid cells, revealing complexities in allele selection during reprogramming.
- By integrating ASE with transcriptomic profiling, it provides a framework for understanding allele-specific regulatory networks in hybrid cellular states.
- The identification of signaling pathways linked to reprogramming efficiency uncovers potential molecular mechanisms limiting placental cell plasticity.
- These insights have broader implications for interspecies stem cell research, including therapeutic applications and understanding genomic conflicts during development.
Conclusion
- Mule-derived iPSCs from different tissue sources vary in reprogramming efficiency, with placental fibroblasts being less efficient.
- Allele-specific expression reveals preferential usage of donkey alleles in key pluripotency genes, especially pronounced in placental fibroblast-derived iPSCs, highlighting the role of genomic imprinting.
- The PI3K-AKT pathway is a candidate mechanism that may influence these differences.
- This comparative study advances understanding of how hybrid genomes regulate cell identity and pluripotency, which is crucial for refining interspecies reprogramming technologies and studying developmental biology in hybrid animals.
Cite This Article
APA
Liu F, Zhang J, Kong L, Wu R, Jiang Q, Lu Y, Li X.
(2025).
Comparative Study of Reprogramming Efficiency and Regulatory Mechanisms of Placental- and Fibroblast-Derived Induced Pluripotent Stem Cells (iPSCs) in Mules.
Curr Issues Mol Biol, 47(8), 671.
https://doi.org/10.3390/cimb47080671 Publication
Researcher Affiliations
- State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- Research Center for Animal Genetic Resources of Mongolia·Plateau, College of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- Inner Mongolia SaiKexing, Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot 011517, China.
- State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- Research Center for Animal Genetic Resources of Mongolia·Plateau, College of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- Liangzhu Laboratory, Zhejiang University Medical Center, Hangzhou 311113, China.
- College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China.
- State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- Research Center for Animal Genetic Resources of Mongolia·Plateau, College of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- National Center of Technology Innovation for Dairy Industry, Hohhot 010020, China.
- State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- Research Center for Animal Genetic Resources of Mongolia·Plateau, College of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- National Center of Technology Innovation for Dairy Industry, Hohhot 010020, China.
- College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China.
- State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- Research Center for Animal Genetic Resources of Mongolia·Plateau, College of Life Sciences, Inner Mongolia University, Hohhot 010070, China.
- Inner Mongolia SaiKexing, Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot 011517, China.
- National Center of Technology Innovation for Dairy Industry, Hohhot 010020, China.
Conflict of Interest Statement
The authors declare no conflicts of interest.
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