000163291 001__ 163291
000163291 005__ 20251024172259.0
000163291 0247_ $$2doi$$a10.1186/s13287-025-04671-1
000163291 0248_ $$2sideral$$a145752
000163291 037__ $$aART-2025-145752
000163291 041__ $$aeng
000163291 100__ $$0(orcid)0000-0001-9818-508X$$aBarrachina, Laura$$uUniversidad de Zaragoza
000163291 245__ $$aGeneration of equine induced pluripotent stem cells from cells of embryonic, perinatal and adult tissues
000163291 260__ $$c2025
000163291 5060_ $$aAccess copy available to the general public$$fUnrestricted
000163291 5203_ $$aBackground: Regenerative therapies are quickly expanding to application in equine patients because of their importance as sporting and companion animals. Furthermore, aligning with a One Health concept, veterinary medicine offers a unique platform for preclinical studies. While mesenchymal stem/stromal cells (MSCs) therapies are already used in treating horses, strategies involving induced pluripotent stem cells (iPSCs) are poorly developed. iPSCs present great potential for therapy and disease modelling, but their consistent generation in horses requires further investigation into the source of somatic cells and the reprogramming method and conditions.
Methods: The reprogramming potential of equine cells from tissues of three developmental origins was compared: prenatal (embryo-derived MSCs, eMSCs), perinatal (cord blood-derived MSCs, CB-MSCs) and adult (articular chondrocytes, ACs). Two reprogramming methods (retroviral, lentiviral) and different culture conditions (serum/serum-free, feeder cells/feeder-free, with/without small molecules) were tested. Pluripotent gene expression was analyzed at different time-points to reveal transcriptomic changes associated with reprogramming. The generated equine iPSCs (eqiPSCs) were characterized by alkaline phosphatase (AP) staining, expression of pluripotent genes and proteins, three-germ layer differentiation (embryoid body) and karyotype.
Results: Using a lentiviral vector with serum-free media and feeder cells resulted in the most favorable conditions for eqiPSCs reprogramming, but adding small molecules had a negative effect. Equine CB-MSCs and ACs were only partially reprogrammed and could not be efficiently expanded in culture. Only eMSCs generated putative eqiPSCs that met the cellular, molecular and functional criteria of pluripotent cells. Equine eMSCs showed higher proliferation and basal expression of pluripotent genes compared to CB-MSCs and ACs, and showed the highest upregulation of pluripotent genes along reprogramming.
Conclusions: The developmental stage of the starting cell strongly influences their reprogramming potential in equine species. This has been suggested for human and other animal species, but direct comparison of equine cells from prenatal, perinatal and adult sources has not been reported before. Novel preliminary insight into the transcriptomic changes of different equine cell types during reprogramming, and on the effect of different culture conditions, can contribute improving the generation of eqiPSCs. While transgene-free methods are the goal, putative eqiPSCs are critical to enlarge our knowledge on animal iPSC biology.
000163291 536__ $$9info:eu-repo/grantAgreement/EC/H2020/101026825/EU/Cartilage derived from equine induced pluripotent stem cells: an in vitro and ex vivo One Medicine approach for osteoarthritis/CAREQiPSC$$9This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 101026825-CAREQiPSC$$9info:eu-repo/grantAgreement/ES/MICINN/PID2020-116352GB-I00
000163291 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttps://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
000163291 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000163291 700__ $$aIvanovska, Ana
000163291 700__ $$aEslami Arshaghi, Tarlan
000163291 700__ $$aO’Brien, Aisling
000163291 700__ $$0(orcid)0000-0002-1481-9805$$aCequier, Alina$$uUniversidad de Zaragoza
000163291 700__ $$aMurphy, Mary
000163291 700__ $$aHollinshead, Fiona
000163291 700__ $$0(orcid)0000-0003-3289-2675$$aRodellar, Clementina$$uUniversidad de Zaragoza
000163291 700__ $$aBarry, Frank
000163291 7102_ $$11001$$2420$$aUniversidad de Zaragoza$$bDpto. Anatom.,Embri.Genét.Ani.$$cÁrea Genética
000163291 7102_ $$11009$$2617$$aUniversidad de Zaragoza$$bDpto. Patología Animal$$cÁrea Medicina y Cirugía Animal
000163291 773__ $$g16, 1 (2025), 547 [18 pp.]$$pStem cell res. ther.$$tStem cell research & therapy$$x1757-6512
000163291 8564_ $$s3271251$$uhttps://zaguan.unizar.es/record/163291/files/texto_completo.pdf$$yVersión publicada
000163291 8564_ $$s2259185$$uhttps://zaguan.unizar.es/record/163291/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000163291 909CO $$ooai:zaguan.unizar.es:163291$$particulos$$pdriver
000163291 951__ $$a2025-10-24-16:56:34
000163291 980__ $$aARTICLE