祖细胞群失调可能导致胎盘发育不良和胎盘功能不全的发病机制。侧群细胞具有祖细胞特性。最近的人类滋养层侧群分离鉴定出 8 个特定基因（CXCL8、ELL2、GATA6、HK2、HLA-DPB1、INTS6、SERPINE3 和 UPP1）的富集（Gamage 等人，2020 年，干​​细胞 Rev Rep）。我们对人胎盘和胎盘功能不全性疾病（先兆子痫和胎儿生长受限（FGR））中的这些滋养层侧群标记进行了表征。在早产对照、先兆子痫和 FGR 胎盘切片中，滋养层侧群标记定位于胎盘绒毛基底膜内衬的单核滋养层（n = 3，3 个标记组/连续切片）。对类器官人滋养层干细胞 (hTSC) 向绒毛外滋养层 (EVT) 分化模型的单细胞转录组学分析（Shannon 等人，2022 年，Development）发现，所有侧群基因均富集于单核滋养层和致力于分化的滋养层中在 hTSC 培养条件下。通过 96 小时 hTSC 分化为 EVT 或合体滋养层 (n = 5) 的体外验证表明，ELL2 和 HK2 随着分化而增加（分别为 p < 0.0024、p < 0.0039）。 CXCL8 和 HLA-DPB1 下调（分别为 p<<0.030、p<0.011）。 GATA6 和 INTS6 仅随着 EVT 分化而增加，UPP1 随着合胞化而减少。 SERPINE3 无法检测到。在人胎盘中测量滋养层侧群标记 mRNA（< 34 周妊娠；n = 78 个先兆子痫，n = 30 个 FGR，n = 18 个妊娠匹配对照）。与对照组相比，先兆子痫（分别为 p = 0.0006、p < 0.0001、p = 0.0335）和 FGR 胎盘（分别为 p = 0.0065、p < 0.0001、p = 0.0001）中 ​​ELL2、HK2 和 CXCL8 升高。在患有先兆子痫和 FGR 的妊娠中，胎盘 GATA6 降低（分别为 p = 0.0014，p = 0.0146）。 仅 FGR 时胎盘 INTS6 降低 (p<<0.0001)。这项研究确定了富含侧群标记的独特滋养层子集的定位。一些侧群标志物的异常表达可能表明胎盘功能不全中独特的滋养层亚型受到破坏。© 2024。作者。

Dysregulated progenitor cell populations may contribute to poor placental development and placental insufficiency pathogenesis. Side-population cells possess progenitor properties. Recent human trophoblast side-population isolation identified enrichment of 8 specific genes (CXCL8, ELL2, GATA6, HK2, HLA-DPB1, INTS6, SERPINE3 and UPP1) (Gamage et al. 2020, Stem Cell Rev Rep). We characterised these trophoblast side-population markers in human placenta and in placental insufficiency disorders: preeclampsia and fetal growth restriction (FGR). Trophoblast side-population markers localised to mononuclear trophoblasts lining the placental villous basement membrane in preterm control, preeclamptic and FGR placental sections (n = 3, panel of 3 markers/serial section). Analysis of single-cell transcriptomics of an organoid human trophoblast stem cell (hTSC) to extravillous trophoblast (EVT) differentiation model (Shannon et al. 2022, Development) identified that all side-population genes were enriched in mononuclear trophoblast and trophoblasts committed to differentiation under hTSC culture conditions. In vitro validation via 96 h time course hTSC differentiation to EVTs or syncytiotrophoblasts (n = 5) demonstrated ELL2 and HK2 increased with differentiation (p < 0.0024, p < 0.0039 respectively). CXCL8 and HLA-DPB1 were downregulated (p < 0.030, p < 0.011 respectively). GATA6 and INTS6 increased with EVT differentiation only, and UPP1 reduced with syncytialisation. SERPINE3 was undetectable. Trophoblast side-population marker mRNA was measured in human placentas (< 34-weeks' gestation; n = 78 preeclampsia, n = 30 FGR, and n = 18 gestation-matched controls). ELL2, HK2 and CXCL8 were elevated in preeclamptic (p = 0.0006, p < 0.0001, p = 0.0335 respectively) and FGR placentas (p = 0.0065, p < 0.0001, p = 0.0001 respectively) versus controls. Placental GATA6 was reduced in pregnancies with preeclampsia and FGR (p = 0.0014, p = 0.0146 respectively). Placental INTS6 was reduced with FGR only (p < 0.0001). This study identified the localisation of a unique trophoblast subset enriched for side-population markers. Aberrant expression of some side-population markers may indicate disruptions to unique trophoblast subtypes in placental insufficiency.© 2024. The Author(s).