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从哥廷根小型猪中分离和鉴定乳腺上皮细胞:与 IMI-ConCePTION 项目的杂种猪细胞的比较研究。

Isolation and characterization of mammary epithelial cells derived from Göttingen Minipigs: A comparative study versus hybrid pig cells from the IMI-ConcePTION Project.

机构信息

Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell 'Emilia, 40064 Bologna, Italy; Health Sciences and Technologies-Interdepartmental Center for Industrial Research (CIRI-SDV), Alma Mater Studiorum-University of Bologna, 40126 Bologna, Italy.

Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell 'Emilia, 40064 Bologna, Italy.

出版信息

Res Vet Sci. 2024 Jun;172:105244. doi: 10.1016/j.rvsc.2024.105244. Epub 2024 Mar 27.

DOI:10.1016/j.rvsc.2024.105244
PMID:38554548
Abstract

The value of pig as "large animal model" is a well-known tool for translational medicine, but it can also be beneficial in studying animal health in a one-health vision. The ConcePTION Project aims to provide new information about the risks associated with medication use during breastfeeding, as this information is not available for most commonly used drugs. In the IMI-Conception context, Göttingen Minipigs have been preferred to hybrid pigs for their genetic stability and microbiological control. For the first time, in the present research, three primary cell cultures of mammary epithelial cells were isolated and characterized from Göttingen Minipigs (mpMECs), including their ability to create the epithelial barrier. In addition, a comparative analysis between Göttingen Minipigs and commercial hybrid pig mammary epithelial cells (pMECs) was conducted. Epithelial markers: CKs, CK18, E-CAD, ZO-1 and OCL, were expressed in both mpMECs and pMECs. RT2 Profiler PCR Array Pig Drug Transporters showed a similar profile in mRNA drug transporters. No difference in energy production under basal metabolic condition was evidenced, while under stressed state, a different metabolic behaviour was shown between mpMECs vs pMECs. TEER measurement and sodium fluorescein transport, indicated that mpMECs were able to create an epithelial barrier, although, this turned out to be less compact than pMECs. By comparing mpMECs with mammary epithelial cells isolated from Hybrid pigs (pMECs), although both cell lines have morphological and phenotypic characteristics that make them both useful in barrier studies, some specific differences exist and must be considered in a translational perspective.

摘要

猪作为“大动物模型”的价值是转化医学的众所周知的工具,但它在研究动物健康的整体健康视野中也可能是有益的。ConCePTION 项目旨在提供与哺乳期使用药物相关的风险的新信息,因为大多数常用药物都没有这方面的信息。在 IMI-Conception 背景下,由于遗传稳定性和微生物控制,哥廷根小型猪被优先于杂种猪用于研究。本研究首次从哥廷根小型猪(mpMECs)中分离并鉴定了三种原代乳腺上皮细胞培养物,包括其形成上皮屏障的能力。此外,还对哥廷根小型猪和商业杂交猪乳腺上皮细胞(pMECs)进行了比较分析。上皮标志物:CKs、CK18、E-CAD、ZO-1 和 OCL 在 mpMECs 和 pMECs 中均有表达。RT2 Profiler PCR 阵列猪药物转运蛋白显示在 mRNA 药物转运蛋白方面具有相似的特征。在基础代谢条件下,未发现能量产生的差异,而在应激状态下,mpMECs 与 pMECs 之间表现出不同的代谢行为。TEER 测量和荧光素钠转运表明,mpMECs 能够形成上皮屏障,尽管其致密程度不如 pMECs。通过将 mpMECs 与从杂种猪中分离的乳腺上皮细胞(pMECs)进行比较,尽管这两种细胞系都具有使它们在屏障研究中都有用的形态和表型特征,但存在一些特定的差异,在转化研究中必须考虑这些差异。

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