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백상기,이인원,이연지,서보경,최정우,김태석,황보철,이준희 사단법인 한국동물생명공학회 2023 Journal of Animal Reproduction and Biotechnology Vol.38 No.4
Background: Porcine pluripotent stem cells (pPSCs) would provide enormous potential for agriculture and biomedicine. However, authentic pPSCs have not established yet because standards for pPSCs-specific markers and culture conditions are not clear. Therefore, the present study reports comparative pluripotency characteristics in porcine induced pluripotent stem cells (piPSCs) derived from different viral transduction and reprogramming factors [Lenti-iPSCs (OSKM), Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM)]. Methods: Porcine fibroblasts were induced into Lenti-iPSCs (OSKM) and Lenti-iPSCs (OSKMNL) by using Lentiviral vector and Sev-iPSCs (OSKM) by using Sendaiviral vector. Expressions of endogenous or exogenous pluripotency-associated genes, surface marker and in vitro differentiation in between Lenti-piPSCs (OSKM), Lenti-iPSCs (OSKMNL) and Sev-piPSCs (OSKM) were compared. Results: Colonial morphology of Lenti-iPSCs (OSKMNL) closely resembles the naïve mouse embryonic stem cells colony for culture, whereas Sev-iPSCs (OSKM) colony is similar to the primed hESCs. Also, the activity of AP shows a distinct different in piPSCs (AP-positive (+) Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM), but AP-negative (-) LentiiPSCs (OSKM)). mRNAs expression of several marker genes (OCT-3/4, NANOG and SOX2) for pluripotency was increased in Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM), but Sev-iPSCs (OSKM). Interestingly, SSEA-1 of surface markers was expressed only in Sev-iPSCs (OSKM), whereas SSEA-4, Tra-1-60 and Tra-1-81 were positively expressed in Lenti-iPSCs (OSKMNL). Exogenous reprogramming factors continuously expressed in Lenti-iPSCs (OSKMNL) for passage 20, whereas Sev-iPSCs (OSKM) did not express any exogenous transcription factors. Finally, only Lenti-iPSCs (OSKMNL) express the three germ layers and primordial germ cells markers in aggregated EBs. Conclusions: These results indicate that the viral transduction system of reprograming factors into porcine differentiated cells display different pluripotency characteristics in piPSCs.
누드 마우스에서 Encephalomycocarditis D 변이종 바이러스의 인슈린 의존성 당뇨병의 유발
백상기,윤지원,신승일 ( Sang Gi Paik,Ji Won Yoon,Seung Il Shin ) 생화학분자생물학회 1986 BMB Reports Vol.19 No.4
The possibility of the host thymus-dependent functions in the development of insulin-dependent diabetes mellitus was investigated in athymic nude (nu/nu) mice and euthymic heterozygous (+/nu) littermates of CD-1 origin, injecting either the multiple subdiabetogenic doses of streptozotocin (STZ) or the D-variant of encephalomyocarditis (EMC-D) virus. A single high dose of STZ caused a rapid hyperglycemia in both +/nu and nu/nu male mice. However, multiple low doses of STZ (30 or 35 ㎎/㎏ b.w./d, for 5 days) induced delayed hyperglycemia only in +/nu male mice ($gt;85%), and not in nu/nu male mice ($lt;17%). It clearly shows inhibitory effect of the nude gene in induction of diabetes in CD-1 mice by multiple low doses of STZ. The EMC-D virus produced hyperglycemia and insulitis in both +/nu and nu/nu male mice (76% diabetic and 73% diabetic, respectively). This shows that the immunological competence of the host may have no effect on diabetogenesis by EMC-D virus. Independent of their thymic function, female mice were quite resistant to the diabetogenic effect of the EMC-D virus. In EMC-D virus infected diabetic mice, the gross necrosis and insulitis were correlated only with the degree of hyperglycemia, regardless of the presence or absence of thymus. These results indicate that diabetes induction by EMC-D virus is not mediated by thymus (T-cell)-dependent processes.