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Derivation of embryonic stem cells from cloned blastocysts using improved somatic cell nuclear transfer in common marmoset

ABSTRACT: The common marmoset (Callithrix jacchus) is a genetically modifiable non-human primate increasingly used in biomedical research. However, methods to preserve or expand valuable genetic resources in this species remain underdeveloped. Here, we established a method for deriving embryonic stem cells (ESCs) from blastocysts generated by somatic cell nuclear transfer (SCNT) in the marmoset. Injection of mRNA encoding the histone demethylase Kdm4d enabled efficient reprogramming of somatic nuclei, allowing blastocyst formation in 14.5% from fibroblasts. However, nuclear transfer ESCs (ntESCs) were not established from these blastocysts. Combining this method with a G9a histone methyltransferase inhibitor (RK-701) improved blastocyst quality and allowed derivation of ntESC lines, including wildtype and GFP-transgenic lines. These ntESCs exhibited normal karyotypes and pluripotency both in vitro and in vivo. Nuclear and mitochondrial DNA analyses confirmed their nuclear donor origin and cytoplasmic inheritance from recipient oocytes. Transcriptome analysis identified abnormally expressed genes in ntESCs present in a line-dependent and -independent manner, suggesting partial reprogramming resistance that may inform future optimization of SCNT in primates. Our study establishes marmoset SCNT method enabling derivation of ntESCs which may provide a new platform for preserving and engineering marmoset genetic resources.

ORGANISM(S): Callithrix jacchus

PROVIDER: GSE305965 | GEO | 2025/09/29

REPOSITORIES: GEO

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