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In vitro induced pluripotency from urine-derived cells in porcine

World Journal of Stem Cells 2022年第3期14卷 231-244页

作者： Kaiana Recchia Lucas Simões Machado Ramon Cesar Botigelli Naira Caroline Godoy Pieri Gabriela Barbosa Raquel Vasconcelos Guimarães de Castro mariana groke marques Laís Vicari de Figueiredo Pessôa Paulo Fantinato Neto Flávio Vieira Meirelles Aline Fernanda de Souza Simone Maria Massami Kitamura Martins Fabiana Fernandes Bressan Department of Surgery Faculty of Veterinary Medicine and Animal SciencesUniversity of São PauloPirassununga 13635-000São PauloBrazil Department of Pharmacology and Biotechnology Institute of BioscienceSão Paulo State UniversityBotucatu 18618-689São PauloBrazil Department of Veterinary Medicine Faculty of Animal Sciences and Food EngineeringUniversity of São PauloPirassununga 13635-000São PauloBrazil Embrapa Suínos e Aves Empresa Brasileira de Pesquisa AgropecuáriaConcordia 89715-899Santa CatarinaBrazil Department of Animal Sciences Faculty of Animal Sciences and Food EngineeringUniversity of São PauloPirassununga 13635-000São PauloBrazil

BACKGROUND The generation of induced pluripotent stem cells(iPSC)has been a game-changer in translational and regenerative medicine;however,their large-scale applicability is still hampered by the scarcity of accessible,safe,and reproducible *** porcine model is a large biomedical model that enables translational applications,including gene editing,long term in vivo and offspring analysis;therefore,suitable for both medicine and animal *** To reprogramme in vitro into pluripotency,and herein urine-derived cells(UDCs)were isolated from porcine *** The UDCs were reprogrammed in vitro using human or murine octamer-binding transcription factor 4(OCT4),SRY-box2(SOX2),Kruppel-like factor 4(KLF4),and C-MYC,and cultured with basic fibroblast growth factor(bFGF)*** characterize the putative porcine iPSCs three clonal lineages were submitted to immunocytochemistry for alkaline phosphatase(AP),OCT4,SOX2,NANOG,TRA181 and SSEA 1 *** transcripts related to the pluripotency(OCT4,SOX2 and NANOG)were analyzed via reverse transcription quantitative realtime polymerase chain reaction in different time points during the culture,and all three lineages formed embryoid bodies(EBs)when cultured in suspension without bFGF *** The UDCs were isolated from swine urine samples and when at passage 2 submitted to in vitro *** of putative iPSCs were obtained only from UDCs transduced with the murine factors(mOSKM),but not from human factors(hOSKM).Three clonal lineages were isolated and further cultured for at least 28 passages,all the lineages were positive for AP detection,the OCT4,SOX2,NANOG markers,albeit the immunocytochemical analysis also revealed heterogeneous phenotypic profiles among lineages and passages for NANOG and SSEA1,similar results were observed in the abundance of the endogenous transcripts related to pluripotent *** the clonal lineages when cultured in suspension without

关键词： Induced pluripotent stem cells Noninvasive Pluripotency Reprogramming Urine Porcine

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Effect of Oocyte Recovery Techniques on in Vitro Production of Swine Embryos

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Open Journal of Animal Sciences 2015年第4期5卷 467-473页

作者： mariana groke marques Flavia Regina Oliveira de Barros Marcelo Demarchi Goissis mariana Ianello Giassetti Mayra Elena Ortiz D’Avila Assumpcao Jose Antonio Visintin EMBRAPA Swine and Poultry Research Center ConcordiaBrazil Department of Animal Reproduction School of Veterinary Medicine and Animal SciencesUniversity of Sao PauloSao PauloBrazil

In Vitro production of swine embryos is a valuable tool to generate clones and genetically modified pigs during a short period of time. However, the efficiency of the existing methods is extremely low and the oocyte quality and quantity represent important obstacles on the success of in Vitro production of embryos. Therefore, the aim of this study was to compare the in Vitro maturation, fertilization and subsequent embryo development rates of oocytes recovered by ovary slicing or follicular aspiration. The oocyte recovery rate (grade 1 COC/ovary) was higher (p = 0.0083) in the slicing group when compared to the aspiration group. No differences were observed between groups regarding in Vitro maturation and early cleavage rates. A higher percentage of oocytes recovered by follicular aspiration reached the blastocyst stage after IVF when compared to the ovary slicing method (p = 0.0395). However, no difference on blastocyst cell number was observed. Although the recovery of oocytes using the slicing technique yielded more grade 1 oocytes per ovary than the aspiration method, the number of oocytes that reached the blastocyst stage after IVF by the slicing method was lower when compared with oocytes obtained by aspiration, as observed by lower blastocyst rates. In conclusion, the follicular aspiration is the method of choice for porcine in Vitro embryo production.

关键词： Oocyte Recovery Oocyte Maturation HSP70 Embryo Development Swine

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