陈老湿最近又迷上一款革命性的细胞成像设备-- Incucyte^® 长时间实时活细胞成像系统，国内这款仪器用得怎么样呢？让陈老湿细细道来！去除预印版和综述，2022年上半年使用Incucyte^® 发表的文章有大几十篇（文献列表见文末），仅第一作者就覆盖了至少40个单位。
 
以下是发表至少2篇以上文章的单位列表：

• 澳门大学中医研究院（4篇）
• 复旦大学上海癌症中心（3篇）
• 中科院动物研究所（3篇）
• 滨州医学院（2篇）
• 郑州大学基础医学院（2篇）
• 中山大学附属第六医院（2篇）

 
可见，很多单位可以利用Incucyte^® 实时、高效地获得细胞信息和可信的研究成果，陈老湿去一些单位拜访的时候，发现这台仪器都是抢着使用的，原因结尾会总结，首先一点就是Incucyte^® 应用非常广泛，从这些文章上看，最多的应用是细胞增殖 & 细胞活性实验，然后是划痕迁移实验，当然还有3D拍摄、神经细胞分析、吞噬等亮点应用。
 
陈老湿就选几篇和大家唠一下：

 
 
Incucyte^® 多点开花应用
山东齐鲁医院
癌症治疗新靶点^【1】

ETS 转录因子 GABPA 长期以来一直被认为是一种致癌因子，最近有人提出由于其对端粒酶活化的关键作用，可以作为癌症治疗的靶点。但 GABPA对细胞肾细胞癌（ccRCC）形成的机理尚不清楚, 利用 siRNA 和表达载体对 GABPA 等因子的表达进行调控发现，GABPA 耗竭虽然抑制端粒酶表达，但显著增强了增殖、侵袭和ccRCC 细胞的干性，而 GABPA 过表达表现出相反的作用，强烈抑制体内转移和致癌。TGFBR2 被鉴定为 GABPA 靶基因，GABPA 通过它控制 TGFβ信号控制ccRCC 表型。Incucyte^® 被用于各种细胞表征实验。


细胞划痕迁移实验：证明GABPA过表达会抑制ccRCC细胞系（A498，786-O）的迁移
  二维细胞增殖实验（左）和三维肿瘤球实验（D,E）表明，沉默GABPA表达加强ccRCC细胞系（A498，786-O）的增殖（D图），GABPA载体增加表达则减弱增殖（E图）。
  
划痕实验，2D和3D拍摄，Incucyte^®都在行！
 
 
Incucyte^®的高通量
中科院动物研究所
干细胞防衰老剂的小分子筛选^【2】

使用 Incucyte^® S3活细胞成像系统，通过监测上千种 FDA 批准的药物对Werner综合征间充质干细胞 （WS hMSCs） 的增殖活性影响，建立了一种高通量药物筛选方法。1,622 种 FDA 批准的药物在一定浓度下持续作用于WS hMSCs 6天。通过初步筛选，确定了前 20 名候选药物。这些候选药物包括卡博替尼、比卡鲁胺、没食子酸 (GA)等。GA 是一种天然酚类化合物，广泛存在于茶、水果和酒中。通过功能性实验，发现GA作为干细胞防衰老剂的活性比二甲双胍和槲皮素还要好。
 

用Incucyte^® 计算药物处理6天后的细胞汇合度的与对照的比值，值越大说明药物增强细胞增殖的能力越强。
 
Incucyte^®一次性可以放6个孔板，其通量可以满足上千个化合物的筛选！
 
 
Incucyte^®吞噬作用
南京中医药大学
天然产物AD药物^【3】

临床证据表明，绝经后女性患阿尔茨海默病 (AD) 的可能性几乎是同龄男性的两倍，雌激素与AD的发生密切相关。雌激素受体ERβ 主要分布在海马体和心血管系统中，ERβ 选择性激动剂是一种有价值的抗神经退行性疾病的药物。在这项研究中，确定了一种天然产物广藿香醇 (PTA) 作为选择性 ERβ激动剂改善雌性 APP/PS1 小鼠的认知缺陷，并探索其潜在机制。小鼠体内实验发现PTA通过促进小胶质细胞的吞噬作用，减少海马中的淀粉样斑块沉积；在体外也观察到 PTA 的治疗效果：PTA (5, 10, 20 μM) 剂量依赖性增加原代小胶质细胞和 BV2 细胞中 o-FAM-Aβ42 的吞噬作用，本研究探讨了选择性 ERβ 激动剂在改善 AD 并突出 PTA 作为抗疾病药物先导化合物的潜力。
  PTA可以增强BV2细胞对FAM标记的Aβ42的吞噬（绿色荧光增多，上面两图以及图d），但是不影响Aβ42的降解（图e）。
 
Incucyte^®有各种配套试剂，可以满足吞噬，内吞，代谢等实验。
 
 

• 培养箱内可长达数周的连续观察，最短几分钟间隔拍摄，减少人力，防止过多操作对细胞的伤害；
• 6个板位，分别独立设置检测程序，可以兼容各种孔板和培养皿，通量高；
• 高效简便的模块化软件设置和数据分析，输出图片、视频、生长曲线等多指标多参数；
• 大于100种优化过的荧光试剂和耗材及详尽的Protocol.

   
 
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-2022年上半年使用Incucyte^® 发表的文章 -
 

1. Myricetin inhibits interferon-γ-induced PD-L1 and IDO1 expression in lung cancer cells.Biochemical Pharmacology 197 (2022) 114940
2. 17β-estradiol rescues the damage of thiazolidinedione on chicken Sertoli cell proliferation via adiponectin.Ecotoxicology and Environmental Safety 233 (2022) 113308
3. 3,30-Diindolylmethane Enhances Fluorouracil Sensitivity via Inhibition of Pyrimidine Metabolism in Colorectal Cancer.Metabolites 2022,12, 410.
4. An optimized method for obtaining clinical-grade specific cell subpopulations from human umbilical cord-derived mesenchymal stem cells.Cell Prolif. 2022;e13300.
5. ATF3 -activated accelerating effect of LINC00941/lncIAPF on fibroblast-to-myofibroblast differentiation by blocking autophagy depending on ELAVL1/HuR in pulmonary fibrosis.AUTOPHAGY https://doi.org/10.1080/15548627.2022.2046448
6. Cholecystectomy promotes colon carcinogenesis by activating the Wnt signaling pathway by increasing the deoxycholic acid level Center.Cell Communication and Signaling 2022, 20(1):71
7. Correction to: Downregulation of fibronectin 1 attenuates ATRA‑induced inhibition of cell migration and invasion in neuroblastoma cells.Molecular and Cellular Biochemistry (2022) 477:1323–1326
8. Cross-species metabolomic analysis identifies uridine as a potent regeneration promoting factor.Cell Discovery ( 2022) 8:6
9. Cytochrome P450 26A1 regulates the clusters and killing activity of NK cells during the peri-implantation period.J Cell Mol Med. 2022;26:2438–2450.
10. DGKZ promotes TGFβ signaling pathway and metastasis in triple-negative breast cancer by suppressing lipid raft-dependent endocytosis of TGFβR2.Cell Death and Disease (2022) 13:105
11. Dysregulated lipid metabolism blunts the sensitivity of cancer cells to EZH2 inhibitor.eBioMedicine 2022;77:103872
12. Emodin targeting the colonic metabolism via PPARγ alleviates UC by inhibiting facultative anaerobe.Phytomedicine 104 (2022) 154106 Available
13. Exogenous iron impairs the anti-cancer effect of ascorbic acid both in vitro and in vivo.Journal of Advanced Research
14. Fluoride exposure induces lysosomal dysfunction unveiled by an integrated transcriptomic and metabolomic study in bone marrow mesenchymal stem cells.Ecotoxicology and Environmental Safety 239 (2022) 113672
15. GABPA‑activated TGFBR2 transcription inhibits aggressiveness but is epigenetically erased by oncometabolites in renal cell carcinoma.J Exp Clin Cancer Res (2022) 41:173
16. Genome-scale CRISPR–Cas9 screen reveals novel regulators of B7-H3 in tumor cells.J Immunother Cancer 2022;10:e004875.
17. Glycopolymer Engineering of the Cell Surface Changes the Single Cell Migratory Direction and Inhibits the Collective Migration of Cancer Cells.ACS Appl. Mater. Interfaces 2022, 14, 4921−4930
18. hnRNPL-activated circANKRD42 back-splicing and circANKRD42-mediated crosstalk of mechanical stiffness and biochemical signal in lung fibrosis.Molecular Therapy Vol. 30 No 6
19. HTR1A Inhibits the Progression of Triple-Negative Breast Cancer via TGF-𝜷 Canonical and Noncanonical Pathways.Adv. Sci. 2022, 9, 2105672
20. Human lysyl‑tRNA synthetase evolves a dynamic structure that can be stabilized by forming complex.Cellular and Molecular Life Sciences (2022) 79:128
21. Industrially synthesized biosafe vaterite hollow CaCO3 for controllable delivery of anticancer drugs.Materials Today Chemistry 24 (2022) 100917
22. Knockdown of Peroxiredoxin V increased the cytotoxicity of non-thermal plasma-treated culture medium to A549 cells.AGING 2022
23. Large-scale chemical screen identifies Gallic acid as a geroprotector for human stem cells.Protein Cell 2022, 13(7):532–539
24. Ligand-Directed Caging Enables the Control of Endogenous DNA Alkyltransferase Activity with Light inside Live Cells.Angew. Chem. Int. Ed. 2022
25. Limiting dilution assay to quantify the self-renewal potential of cancer stem cells in hepatocellular carcinoma.Methods in Cell Biology, ISSN 0091-679X, https://doi.org/10.1016/bs.mcb.2022.04.010
26. Long non‑coding RNA LSAMP‑1 is down‑regulated in non‑small cell lung cancer and predicts a poor prognosis.Cancer Cell International (2022) 22:181
27. Low doses of niclosamide and quinacrine combination yields synergistic effect in melanoma via activating autophagy-mediated p53-dependent apoptosis.Translational Oncology 21 (2022) 101425
28. Low-dose chloroquine treatment extends the lifespan of aged rats.Protein Cell 2022, 13(6):454–461
29. Mutations and clinical significance of calcium voltage-gated channel subunit alpha 1E (CACNA1E) in non-small cell lung cancer.https://doi.org/10.1016/j.annonc.2022.02.208
30. Network pharmacology and in vitro studies reveal the pharmacological effects and molecular mechanisms of Shenzhi Jiannao prescription against vascular dementia.BMC Complementary Medicine and Therapies (2022) 22:33
31. Nonmetal Graphdiyne Nanozyme-Based Ferroptosis−Apoptosis Strategy for Colon Cancer Therapy.ACS Appl. Mater. Interfaces 2022, 14, 27720−27732
32. Novel prognostic marker LINC00205 promotes tumorigenesis and metastasis by competitively suppressing miRNA-26a in gastric cancer.Cell Death Discovery (2022) 8:5
33. Patchouli alcohol as a selective estrogen receptor β agonist ameliorates AD-like pathology of APP/PS1 model mice.Acta Pharmacologica Sinica (2022) 0:1–16
34. Promoting role of pentraxin-3 in esophageal squamous cell carcinoma.Molecular Therapy
35. Psoralidin, a natural compound from Psoralea corylifolia, induces oxidative damage mediated apoptosis in colon cancer cells.J Biochem Mol Toxicol. 2022;e23051
36. Receptor activity‐modifying protein 1 regulates mouse skin fibroblast proliferation via the Gαi3‑PKA‑CREB‑YAP axis.Cell Communication and Signaling (2022) 20:52
37. Redirecting anti-Vaccinia virus T cell immunity for cancer treatment by AAV-mediated delivery of the VV B8R gene.Molecular Therapy
38. Research on selective uptake of photosensitizer C3N4@RP by different cancer cells.Mater. Res. Express 9 (2022) 065402
39. RNF4 silencing induces cell growth arrest and DNA damage by promoting nuclear targeting of p62 in hepatocellular carcinoma.Oncogene (2022) 41:2275–2286
40. S100A9-CXCL12 activation in BRCA1-mutant breast cancer promotes an immunosuppressive microenvironment associated with resistance to immunotherapy.NATURE COMMUNICATIONS | (2022) 13:1481
41. Schlafen 5 suppresses human immunodeficiency virus type 1 transcription by commandeering cellular epigenetic machinery.Nucleic Acids Research, 2022
42. Shuxuetong injection and its peptides enhance angiogenesis after hindlimb ischemia by activating the MYPT1/LIMK1/Cofilin pathway.Journal of Ethnopharmacology 292 (2022) 115166
43. SIK2 promotes ovarian cancer cell motility and metastasis by phosphorylating MYLK.Molecular Oncology 16 (2022) 2558–2574
44. SLC1A1-mediated cellular and mitochondrial influx of R-2-hydroxyglutarate in vascular endothelial cells promotes tumor angiogenesis in IDH1-mutant solid tumors.Cell Research (2022) 32:638–658
45. SLC35E2 promoter mutation as a prognostic marker of esophageal squamous cell carcinoma.Life Sciences 296 (2022) 120447
46. Suppression of toxicity of the mutant huntingtin protein by its interacting compound, desonide.PNAS 119 (10) e2114303119
47. Systematic Survey of the Regulatory Networks of the Long Noncoding RNA BANCR in Cervical Cancer Cells.J. Proteome Res. 2022, 21, 1137−1152
48. The early‑stage triple‑negative breast cancer landscape derives a novel prognostic signature and therapeutic target.Breast Cancer Research and Treatment (2022) 193:319–330
49. The SF3B1R625H mutation promotes prolactinoma tumor progression through aberrant splicing of DLG1.J Exp Clin Cancer Res (2022) 41:26
50. The two‑faced role of ATF2 on cisplatin response in gastric cancer depends on p53 context.Cell & Bioscience (2022) 12:77
51. Toosendanin, a late‑stage autophagy inhibitor, sensitizes triple‑negative breast cancer to irinotecan chemotherapy.Chinese Medicine (2022) 17:55
52. TSG-6 promotes Cancer Cell aggressiveness in a CD44-Dependent Manner and Reprograms Normal Fibroblasts to create a Pro-metastatic Microenvironment in Colorectal Cancer.Int. J. Biol. Sci. 2022, Vol. 18
53. Upregulation of ADAMDEC1 correlates with tumor progression and predicts poor prognosis in non-small cell lung cancer (NSCLC) via the PI3K/AKT pathway.Thorac Cancer. 2022;13:1027–1039

-参考文献-
【1】GABPA‑activated TGFBR2 transcription inhibits aggressiveness but is epigenetically erased by oncometabolites in renal cell carcinoma.J Exp Clin Cancer Res (2022) 41:173
【2】Large-scale chemical screen identifies Gallic acid as a geroprotector for human stem cells.Protein Cell 2022, 13(7):532–539
【3】Patchouli alcohol as a selective estrogen receptor β agonist ameliorates AD-like pathology of APP/PS1 model mice.Acta Pharmacologica Sinica (2022) 0:1–16