Rapid generation of marsupial induced pluripotent stem cells suitable for use in next-generation conservation

Fat-tailed dunnart iPSCs generated from neonatal fibroblasts integrate into both inner cell mass and trophoblast after injection into 8-cell mouse blastocysts.

• Why it matters: Conservation work in threatened marsupials needs faster, flexible platforms for genetic selection or modification, but transgene-free iPSC derivation can be time consuming and costly.
• What they did: Neonatal Sminthopsis crassicaudata fibroblasts were reprogrammed with doxycycline-inducible Oct4, Sox2, Klf4, Lin28 and Myc, then characterized by transcriptomics, epigenetics, trilineage differentiation, embryoid body formation, and mouse blastocyst injection.
• The result: The resulting iPSC lines showed strong activation of pluripotency-associated factors (including POU5F1, POU5F3, SALL4) and differentiated into all three germ layers, supporting rapid iPSC-based conservation technology development.

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