用于识别iPSC的最常见基因是TRA-1-60、阶段特异性胚胎抗原4(SSEA4)和TRA-1-81。SSEA-4表达似乎先于TRA-1-60和TRA-1-80的表达,它们只能在后期分化阶段检测到。TRA-1-60和TRA-1-80是糖蛋白鬼萼素的独特表位,用于鉴定和分离ESC。 靶向治疗性细胞治疗...
应用实例:在iPSCs的纯化过程中,通过流式细胞术筛选出表达高水平SSEA-4和TRA-1-60的细胞群体,可以显著提高细胞纯度,确保下游实验的可靠性。此外,免疫荧光染色可用于确认细胞表面标志物的表达模式,进一步验证细胞的多能性状态。 优势与局限: 优势:能够快速、准确地检测和定量细胞表面标志物,适用于大规模细胞筛选与纯化...
iPSC pool purification scheme using Magnetic Activated Cell Sorting (MACS) of TRA-1-60 or SSEA4 expressing cells.Wenli, YangYing, LiuKatherine, J. SlovikJoseph, C. WuStephen, A. DuncanDaniel, J. RaderEdward, E. Morrisey
- 表达胚胎干细胞表面标记物,如SSEA-3、SSEA-4、TRA-1-60和TRA-1-81等 - 缺乏体细胞特异性标记物的表达 5. 细胞形态的可变性: - iPSCs在不同的培养条件下形态略有差异 - 当iPSCs分化时,细胞形态会发生相应变化 准确评估iPSCs的形态非常重要,因为它反映了细胞的干细胞状态和多能性。形态学观察结合分子...
1 panel C Quantitative FLOW cytometry analysis COL6.4uOCT3/4: 99.0 %SSEA: 97.7 %TRA-1–60: 92.7 %COL6.22cOCT3/4: 98.3 %SSEA: 91.9 %TRA-1–60: 94.1 % Fig. 1 panel D Genotype Infinium GSA-24 v2.0 SNP array (Illumina) Arr(1–22,X)x2 Supplementary Fig. 1 and Supplementary ...
1. 准备EB形成培养基(见表1)。 2.按照以下方法,制备EB接种培养基:30 µL 5 mM Y-27632加入15 mL EB形成培养基中,使其终浓度为10 μM。 3.显微镜下观察确定hPSC培养分化区域。通过刮擦去除分化的区域。 4.将hPSC培养孔中的培养基吸去,用1 mL无菌的PBS洗涤培养孔。
All generated iPSCs showed human embryonic stem cell (hESC)-like morphology and expressed alkaline phosphatase and the pluripotency markers SSEA4, TRA-1-81, TRA-1-60, OCT4, NANOG and LIN28 (Supplementary Figs 1 and 2a). iPSCs were genetically matched with their parental cells by PCR-based...
Stem Cell Research & Therapy (2023) 14:154 A) Page 7 of 24 B) Patient-derived PBMC Reprogrammed iPSC C) iPSC Colonies ALP Staining 100 m 100 m D) Isotype Control iPSC E) Oct4 Tra-1-60 (0.1%) Tra-1-60/ SSEA4 (0.00%) Tra-1-60 (0.1%) Tra-1-60/ SSEA4 (99.9%) SSEA4 ...
TRA-1-60 (PE) Miltenyi Biotec Cat# 130-100-347; RRID: AB_2654227 REA Isotype Control (I)-PE Miltenyi Biotec Cat# 130-104-613; RRID: AB_2661678 Beta Actin Abcam Cat# ab49900; (RRID: AB_867494 Anti-Mouse IgG (HRP) Santa Cruz Cat# sc-2005; RRID: AB_631736 Anti-Mouse IgG (HRP...
1 panel F Quantitative analysis (Flow cytometry) Both lines are positive for cell surface markers TRA-1-60 (DS1.1 99.17%; DS3.1 99.85%) and SSEA4 (DS1.1 99.9%; DS3.1 96.1%) Fig. 1 panel B Genotype Karyotype (G-banding) DS1.1 47,XY,+21, P24DS3.1 47,XY,+21, P22 Resolution 400...