HLA类我表达在人类胎盘和体外模型人滋养层。(一)对于妊娠前三个月的锚定绒毛的插图。HLA-null绒毛细胞(SCT, VCT和细胞的基列(CCC)]是绿色的;HLA-C, E, G + EVT紫色;和抗原+ B + C +和HLA-G−nontrophoblast细胞绒毛的核心是蓝色的。(B)免疫组织化学染色对HLA分子类我acetone-fixed对于妊娠前三个月胎盘部分。这里,pan-class我抗体W6/32(我分子结合所有HLA类)污渍EVT的绒毛核心(nontrophoblast)和细胞列但不是VCT或SCT。染色G233,专门针对HLA-G,增加细胞远离绒毛细胞列,而整个绒毛仍为负值。(C) HLA概要文件(使用W6/32和MEMG-9 HLA-G-specific抗体)的细胞株用作HLA控制表达式通过流式细胞仪:JEG-3(控制HLA的extravillous滋养层);JAR(控制绒毛滋养层); and 2102Ep (nontrophoblast control). (D) FACS analysis of W6/32 and MEMG-9 in JAR, JEG-3, TOs and TSCs. TOs have the profile of villous trophoblast (W6/32−/MEMG-9−; n=4), whereas TSCs have neither villous nor extravillous profiles [W6/32+/MEMG-9−; n=5]. (E) FACS analysis of TSCs grown under proliferative conditions and when differentiated to EVT (n=2). Allele-specific antibodies were used to assess HLA-A (BB7.2) and HLA-B (Bw6) expression in a HLA-genotyped TSC line (BTS5). Undifferentiated TSCs express HLA-A and HLA-B, which is maintained after EVT differentiation. (F) FACS analysis demonstrating upregulation of HLA-G in TSCs following EVT differentiation (n=3). (G) Live staining for W6/32-Alexa488 on TSCs differentiated to either EVT or SCT (n=2). Distinct membrane staining is seen when differentiated to EVT and is absent in SCT. (H) Experimental set-up of the different trophoblast culture conditions. (I) FACS analysis of TSCs grown in 2D versus 3D (passaged more than six times in 3D, n=4) with W6/32 and MEMG-9. The number of cells that are W6/32+/MEMG-9− significantly decreases when cultured in 3D. (J) Quantification of the percentage of cells in the W6/32+/MEMG-9− quadrant under 2D or 3D conditions (data are mean±s.e.m., paired two-tailed Student's t-test, **P=0.0019). Scale bars: 50 µm in G; 150 µm in B. Credit: DOI: 10.1242/dev.199749
