Yang Liheng, Liang Pengfei, Yang Huanghe, Coyne Carolyn B
Department of Integrative Immunobiology, Duke University Medical Center, Durham, NC, USA.
Department of Biochemistry, Duke University Medical Center, Durham, NC, USA.
bioRxiv. 2023 Jul 17:2023.01.12.523752. doi: 10.1101/2023.01.12.523752.
Human trophoblast organoids (TOs) are a three-dimensional culture model that can be used to study various aspects of placental development, physiology, and pathology. Previously, we showed that TOs derived from full-term human placental tissue could be used as models of trophoblast innate immune signaling and teratogenic virus infections. Here, we developed a method to culture TOs under conditions that recapitulate the cellular orientation of chorionic villi , with the multi-nucleated syncytiotrophoblast (STB) localized to the outer surface of organoids and the proliferative cytotrophoblasts (CTBs) located on the inner surface. We show that standard TOs containing the STB layer inside the organoid (STB ) develop into organoids containing the STB on the outer surface (STB ) when cultured in suspension with gentle agitation. STB organoids secrete higher levels of select STB-associated hormones and cytokines, including human chorionic gonadotropin (hCG) and interferon (IFN)-λ2. Using membrane capacitance measurements, we also show that the outermost surface of STB organoids contain large syncytia comprised of >50 nuclei compared to STB organoids that contain small syncytia (<10 nuclei) and mononuclear cells. The growth of TOs under conditions that mimic the cellular orientation of chorionic villi thus allows for the study of a variety of aspects of placental biology under physiological conditions.
人滋养层类器官（TOs）是一种三维培养模型，可用于研究胎盘发育、生理学和病理学的各个方面。此前，我们表明，源自足月人胎盘组织的TOs可作为滋养层固有免疫信号传导和致畸病毒感染的模型。在这里，我们开发了一种在模拟绒毛膜绒毛细胞取向的条件下培养TOs的方法，多核合体滋养层（STB）定位于类器官的外表面，增殖性细胞滋养层（CTB）位于内表面。我们发现，当在温和搅拌下悬浮培养时，类器官内部含有STB层的标准TOs（STB⁻）会发育成外表面含有STB的类器官（STB⁺）。STB⁺类器官分泌更高水平的特定STB相关激素和细胞因子，包括人绒毛膜促性腺激素（hCG）和干扰素（IFN）-λ2。通过膜电容测量，我们还表明，与含有小合体（<10个核）和单核细胞的STB⁻类器官相比，STB⁺类器官的最外表面含有由>50个核组成的大合体。因此，在模拟绒毛膜绒毛细胞取向的条件下培养TOs能够在生理条件下研究胎盘生物学的各个方面。
