Pitrone Maria, Pizzolanti Giuseppe, Tomasello Laura, Coppola Antonina, Morini Lorenzo, Pantuso Gianni, Ficarella Romina, Guarnotta Valentina, Perrini Sebastio, Giorgino Francesco, Giordano Carla
Aldo Galluzzo Laboratory of Regenerative Medicine, Section of Endocrinology, Diabetology and Metabolism, Di.Bi.M.I.S, University of Palermo, 90127 Palermo, Italy.
Oncology Surgery, University of Palermo, 90127 Palermo, Italy.
Int J Mol Sci. 2017 May 23;18(6):1107. doi: 10.3390/ijms18061107.
The stromal vascular cell fraction (SVF) of visceral and subcutaneous adipose tissue (VAT and SAT) has increasingly come into focus in stem cell research, since these compartments represent a rich source of multipotent adipose-derived stem cells (ASCs). ASCs exhibit a self-renewal potential and differentiation capacity. Our aim was to study the different expression of the embryonic stem cell markers (homeobox protein NANOG), (SRY (sex determining region Y)-box 2) and (octamer-binding transcription factor 4) and to evaluate if there exists a hierarchal role in this network in ASCs derived from both SAT and VAT. ASCs were isolated from SAT and VAT biopsies of 72 consenting patients (23 men, 47 women; age 45 ± 10; BMI between 25 ± 5 and 30 ± 5 range) undergoing elective open-abdominal surgery. Sphere-forming capability was evaluated by plating cells in low adhesion plastic. Stem cell markers CD90, CD105, CD29, CD31, CD45 and CD146 were analyzed by flow cytometry, and the stem cell transcription factors NANOG, SOX2 and OCT4 were detected by immunoblotting and real-time PCR. , and interplay was explored by gene silencing. ASCs from VAT and SAT confirmed their mesenchymal stem cell (MSC) phenotype expressing the specific MSC markers CD90, CD105, NANOG, SOX2 and OCT4. silencing induced a significant (70 ± 0.05%) and (75 ± 0.03%) downregulation, whereas silencing did not affect gene expression. Adipose tissue is an important source of MSC, and siRNA experiments endorse a hierarchical role of in the complex transcription network that regulates pluripotency.
内脏和皮下脂肪组织(VAT和SAT)的基质血管细胞组分(SVF)在干细胞研究中越来越受到关注,因为这些区域代表了多能脂肪来源干细胞(ASC)的丰富来源。ASC具有自我更新潜力和分化能力。我们的目的是研究胚胎干细胞标志物(同源盒蛋白NANOG)、(SRY(性别决定区Y)-盒2)和(八聚体结合转录因子4)的不同表达,并评估在源自SAT和VAT的ASC中该网络是否存在层次作用。从72例接受择期开放性腹部手术的自愿患者(23名男性,47名女性;年龄45±10岁;BMI在25±5至30±5范围内)的SAT和VAT活检组织中分离出ASC。通过将细胞接种在低粘附性塑料培养皿中来评估成球能力。通过流式细胞术分析干细胞标志物CD90、CD105、CD29、CD31、CD45和CD146,并通过免疫印迹和实时PCR检测干细胞转录因子NANOG、SOX2和OCT4。通过基因沉默探索、和的相互作用。来自VAT和SAT的ASC证实了它们的间充质干细胞(MSC)表型,表达特异性MSC标志物CD90、CD105、NANOG、SOX2和OCT4。沉默诱导了显著的(70±0.05%)和(75±0.03%)下调,而沉默不影响基因表达。脂肪组织是MSC的重要来源,并且siRNA实验支持在调节多能性的复杂转录网络中的层次作用。