人白色脂肪组织中永生化CD55+祖细胞的成脂特性。

IF 3.5 4区 生物学 Q2 ENDOCRINOLOGY & METABOLISM Adipocyte Pub Date : 2023-11-20 DOI:10.1080/21623945.2023.2283213
Morgane Couchet, Hui Gao, Felix Klingelhuber, Jutta Jalkanen, Thais De Castro Barbosa, Muhmmad Omar-Hmeadi, Lucas Massier, Natalie Krahmer, Niklas Mejhert, Mikael Rydén
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引用次数: 0

摘要

背景:成熟脂肪细胞是出了名的难以离体研究,因此开发了替代的细胞培养系统。最常见的模型之一是人类脂肪祖细胞(hAPCs)。不幸的是,在长时间的培养条件下,这些细胞表现出复制性衰老,这限制了它们在机制研究中的应用。方法:将人端粒酶逆转录酶(TERT)敲入来自腹腔皮下脂肪组织的CD55+ hAPCs的AAVS1位点,并对细胞分化为脂肪细胞前后的细胞进行表征。结果:永生化的TERT-hAPCs保持了与早期传代野生型hAPCs相当的增殖和成脂能力。与此一致,我们的整合转录组学和蛋白质组学分析显示,与野生型hapc相比,tert - hapc表现出强大的脂肪细胞表达谱。脂质转换的功能分析证实了这一点,其中TERT-hAPCs对胰岛素和促脂解刺激(如异丙肾上腺素、二丁基cAMP和肿瘤坏死因子α)表现出明显的反应。此外,TERT-hAPCs可以很容易地在标准的2D和3d培养中培养,蛋白质组学分析显示球形培养条件可以促进脂肪形成。结论:通过描述性和功能性研究,我们证明了人CD55+ hAPCs的永生化是可行的,并且在多次传代中产生的细胞具有稳定的增殖和成脂能力。由于这些细胞是可低温保存的,它们比原代细胞提供了额外的优势,允许在具有相同遗传背景的2D和3D模型系统中重复研究。(234/250)。
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Adipogenic characterization of immortalized CD55+ progenitor cells from human white adipose tissue.

Background: Mature adipocytes are notoriously difficult to study ex vivo and alternative cell culture systems have therefore been developed. One of the most common models are human adipose progenitor cells (hAPCs). Unfortunately, these display replicative senescence after prolonged culture conditions, which limits their use in mechanistic studies.

Methods: Herein, we knocked in human telomerase reverse transcriptase (TERT) into the AAVS1 locus of CD55+ hAPCs derived from abdominal subcutaneous adipose tissue and characterized the cells before and after differentiation into adipocytes.

Results: Immortalized TERT-hAPCs retained proliferative and adipogenic capacities comparable to those of early-passage wild type hAPCs for > 80 passages. In line with this, our integrative transcriptomic and proteomic analyses revealed that TERT-hAPCs displayed robust adipocyte expression profiles in comparison to wild type hAPCs. This was confirmed by functional analyses of lipid turnover where TERT-hAPCs exhibited pronounced responses to insulin and pro-lipolytic stimuli such as isoprenaline, dibutyrul cAMP and tumour necrosis factor alpha. In addition, TERT-hAPCs could be readily cultured in both standard 2D and 3D-cultures and proteomic analyses revealed that the spheroid culture conditions improved adipogenesis.

Conclusion: Through descriptive and functional studies, we demonstrate that immortalization of human CD55+ hAPCs is feasible and results in cells with stable proliferative and adipogenic capacities over multiple passages. As these cells are cryopreservable, they provide the additional advantage over primary cells of allowing repeated studies in both 2D and 3D model systems with the same genetic background. (234/250).

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来源期刊
Adipocyte
Adipocyte Medicine-Histology
CiteScore
6.50
自引率
3.00%
发文量
46
审稿时长
32 weeks
期刊介绍: Adipocyte recognizes that the adipose tissue is the largest endocrine organ in the body, and explores the link between dysfunctional adipose tissue and the growing number of chronic diseases including diabetes, hypertension, cardiovascular disease and cancer. Historically, the primary function of the adipose tissue was limited to energy storage and thermoregulation. However, a plethora of research over the past 3 decades has recognized the dynamic role of the adipose tissue and its contribution to a variety of physiological processes including reproduction, angiogenesis, apoptosis, inflammation, blood pressure, coagulation, fibrinolysis, immunity and general metabolic homeostasis. The field of Adipose Tissue research has grown tremendously, and Adipocyte is the first international peer-reviewed journal of its kind providing a multi-disciplinary forum for research focusing exclusively on all aspects of adipose tissue physiology and pathophysiology. Adipocyte accepts high-profile submissions in basic, translational and clinical research.
期刊最新文献
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