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用于研究睾丸支持细胞的非人灵长类动物模型的特征-与人类睾丸支持细胞的比较。

Characterization of a non-human primate model for the study of testicular peritubular cells-comparison with human testicular peritubular cells.

机构信息

Cell Biology-Anatomy III, Biomedical Center Munich (BMC), Ludwig-Maximilians-Universität München, Großhaderner Strasse 9, Martinsried, Germany.

Laboratory for Functional Genome Analysis LAFUGA, Gene Center, LMU München, Fedor-Lynen-Strasse 25, Munich, Germany.

出版信息

Mol Hum Reprod. 2018 Aug 1;24(8):401-410. doi: 10.1093/molehr/gay025.

Abstract

STUDY QUESTION

Are monkey testicular peritubular cells (MKTPCs) from the common marmoset monkey (Callithrix jacchus) a suitable translational model for the study of human testicular peritubular cells (HTPCs)?

SUMMARY ANSWER

MKTPCs can be isolated and propagated in vitro, retain characteristic markers for testicular peritubular cells and their proteome strongly (correlation coefficient of 0.78) overlaps with the proteome of HTPCs.

WHAT IS KNOWN ALREADY

Smooth-muscle-like peritubular cells form the wall of seminiferous tubules, transport sperm, are immunologically active, secrete a plethora of factors and may contribute to the spermatogonial stem cell niche. Mechanistic studies are hampered by heterogeneity of human samples.

STUDY DESIGN, SIZE, DURATION: We established a culture method for MKTPCs and characterized these cells from six young adult animals (2-3 years). To examine whether they qualify as a translational model we also examined HTPCs from seven men and compared the proteomes of both groups.

PARTICIPANTS/MATERIALS, SETTING, METHODS: We used explant cultures to obtain MKTPCs, which express smooth muscle markers (calponin (CNN1), smooth muscle actin (ACTA2)), lack FSH-receptors (FSHR) and LH-receptors (LHCGR), but possess androgen receptors (AR). MKTPCs can be passaged at least up to eight times, without discernable phenotypic changes. Mass-spectrometry-based analyses of the MKTPC and HTPC proteomes were performed.

MAIN RESULTS AND THE ROLE OF CHANCE

We established a method for isolation and cultivation of MKTPCs, and provide a comprehensive analysis of their protein repertoire. The results let us conclude that MKTPCs are suitable as a non-human primate model to study peritubular cell functions.

LARGE SCALE DATA

List of identified proteins in MKTPCs by liquid chromatography-tandem mass spectrometry is accessible at the ProteomeXchange (identifier PXD009394).

LIMITATIONS, REASON FOR CAUTION: This is an in vitro cellular non-human primate model used to provide a window into the role of these cells in the human testis.

WIDER IMPLICATIONS OF THE FINDINGS

Previous studies with HTPCs from patients revealed a degree of heterogeneity, possibly due to age, lifestyle and medical history of the individual human donors. We anticipate that the new translational model, derived from young healthy non-human primates, may allow us to circumvent these issues and may lead to a better understanding of the role of peritubular cells.

STUDY FUNDING AND COMPETION OF INTEREST(S): This work was supported by grants from the Deutsche Forschungsgemeinschaft (MA 1080/27-1; AR 362/9-1; BE 2296/8-1). The authors declare no competing financial interests.

摘要

研究问题

来自普通狨猴(Callithrix jacchus)的睾丸支持细胞(MKTPC)是否是研究人类睾丸支持细胞(HTPC)的合适转化模型?

总结答案

可以从六只年轻成年动物(2-3 岁)中分离和培养 MKTPC,并保留其睾丸支持细胞的特征标志物,其蛋白质组与 HTPC 的蛋白质组强烈重叠(相关系数为 0.78)。

已知内容

平滑肌样支持细胞形成生精小管的壁,运输精子,具有免疫活性,分泌大量因子,并可能有助于精原干细胞龛。由于人类样本的异质性,机制研究受到阻碍。

研究设计、大小、持续时间:我们建立了 MKTPC 的培养方法,并从六只年轻成年动物(2-3 岁)中对这些细胞进行了特征描述。为了检验它们是否符合转化模型的标准,我们还检查了来自七个男性的 HTPC,并比较了两组的蛋白质组。

参与者/材料、设置、方法:我们使用外植体培养法获得 MKTPC,其表达平滑肌标志物(钙调蛋白 1(CNN1)、平滑肌肌动蛋白(ACTA2)),缺乏促卵泡激素受体(FSHR)和促黄体激素受体(LHCGR),但具有雄激素受体(AR)。MKTPC 至少可以传代 8 次,而没有明显的表型变化。对 MKTPC 和 HTPC 蛋白质组进行基于质谱的分析。

主要结果和机会的作用

我们建立了分离和培养 MKTPC 的方法,并对其蛋白质组进行了全面分析。结果使我们得出结论,MKTPC 适合作为非人类灵长类动物模型来研究支持细胞的功能。

大规模数据

通过液相色谱-串联质谱鉴定的 MKTPC 中的蛋白质列表可在 ProteomeXchange(标识符 PXD009394)上获得。

局限性、谨慎的原因:这是一种体外细胞非人类灵长类动物模型,用于提供人类睾丸中这些细胞作用的窗口。

研究结果的更广泛意义

先前使用来自患者的 HTPC 的研究显示出一定程度的异质性,可能是由于个体人类供体的年龄、生活方式和病史所致。我们预计,来自年轻健康的非人类灵长类动物的新转化模型可能使我们能够避免这些问题,并可能导致对支持细胞作用的更好理解。

研究资金和利益冲突

这项工作得到了德国研究基金会(MA 1080/27-1;AR 362/9-1;BE 2296/8-1)的资助。作者没有竞争利益。

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