Non-heat-stressed Method to Isolate Hepatic Stellate Cells From Highly Steatotic Tumor-bearing Liver Using CD49a.
Non-heat-stressed Method to Isolate Hepatic Stellate Cells From Highly Steatotic Tumor-bearing Liver Using CD49a.
复制标题
使用CD49A将肝脏星状细胞与高度脂肪酸肿瘤的肝脏分离的非热压力方法。
DOI：
10.1016/j.jcmgh.2022.07.006
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发表时间：
2022
影响因子：
7.2
通讯作者：
Ohtani, Naoko
中科院分区：
文献类型：
--
作者：
Cheng, Yi;Yamagishi, Ryota;Nonaka, Yoshiki;Sato-Matsubara, Misako;Kawada, Norifumi;Ohtani, Naoko
随着肥胖的流行，非病毒性、非酒精性脂肪性肝炎相关肝癌的患者数量一直在增加。越来越多的证据表明，肿瘤微环境（TME）对肿瘤进展至关重要。我们以前发现，肝星状细胞（HSC）通过分泌衰老相关的分泌因子抑制抗肿瘤免疫，在肥胖相关肝细胞癌（HCC）的致瘤性TME中发挥关键作用。1，2，3单细胞RNA测序（scRNA-seq）技术的最新进展使得能够表征各种组织中的不同细胞类型。因此，开发一种精确的方法来表征在各种条件下TME中的HSC是必要的，以了解它们在HCC进展中的作用。已经报道了使用逆行方法通过下腔静脉（IVC）灌注从正常肝脏分离HSC。4然而，该方法不适用于肥胖小鼠中的高度脂肪变性肝脏或脂肪性肝炎肿瘤组织，因为肥胖小鼠中脂肪组织的积累阻碍了IVC可视化，从而使得经由IVC的插管困难。因此，我们尝试通过门静脉（PV）分离HSC，这在所有条件下都清晰可见。我们的目的是开发一种在低温（6 ℃）下分离HSC的方法，以最小化酶的过度消化和解离后（37 ℃）的热相关应激，据报道，这会诱导基因如Fos和Jun.5。此外，我们鉴定了在HSC中大量表达的细胞表面标记物，其可用于从鼠和人肝肿瘤组织中分选HSC细胞。
The number of patients with non-viral, nonalcoholic steatohepatitis-associated liver cancer has been increasing with the prevalence of obesity. Increasing evidence shows that the tumor microenvironment (TME) is crucial for tumor progression. We previously showed that hepatic stellate cells (HSCs) play key roles in tumorigenic TME of obesity-associated hepatocellular carcinoma (HCC) by secreting senescence-associated secretory factors that suppress anti-tumor immunity. 1, 2, 3 Recent advances in single-cell RNA-sequencing (scRNA-seq) techniques have enabled the characterization of different cell types in various tissues. Therefore, development of a refined method to characterize HSCs in TME under various conditions is necessary to understand their role in HCC progression.Isolation of HSCs from normal liver via perfusion from the inferior vena cava (IVC) using a retrograde approach has been reported. 4 However, this method is not applicable to highly steatotic liver or steatohepatitic tumor tissues in obese mice as the accumulation of adipose tissue in obese mice hinders IVC visualization, thereby making cannulation via IVC difficult. Therefore, we attempted HSC isolation through the portal vein (PV), which is clearly visible under all conditions. We aimed to develop a method to isolate HSCs at a low temperature (6 C) to minimize over digestion by enzymes and heat-associated stress after dissociation (at 37 C), which reportedly induce genes such as Fos and Jun. 5 Furthermore, we identified a cell surface marker abundantly expressed in HSCs, which is useful for HSC cell sorting from murine and human liver tumor tissues.
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影响因子：
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