HLA I类在人胎盘和体外滋养层模型中的表达。(A)孕早期绒毛锚定示意图。无hla绒毛细胞[SCT, VCT和细胞柱基底(CCC)]为绿色;HLA-C、E和G+ EVT为紫色;绒毛核的HLA-A+、B+、C+和HLA-G−非滋养细胞为蓝色。(B)在丙酮固定的孕早期胎盘切片上进行HLA I类分子免疫组化染色。图中，泛I类抗体W6/32(结合所有HLA I类分子)染色于细胞柱中的绒毛核(非滋养层)和EVT，但不染色于VCT或SCT。用HLA-G特异性的G233染色，随着细胞从绒毛转移到细胞柱而增加，而整个绒毛保持阴性。(C) HLA谱(使用W6/32和MEMG-9，一种HLA- g特异性抗体):JEG-3(细胞外滋养层HLA谱对照);JAR(绒毛滋养层对照); 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
