Interaction between human placental microvascular endothelial cells and a model of human trophoblasts: effects on growth cycle and angiogenic profile.
Interaction between human placental microvascular endothelial cells and a model of human trophoblasts: effects on growth cycle and angiogenic profile.
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DOI：
10.1002/phy2.244
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发表时间：
2014
影响因子：
2.5
通讯作者：
Jones, Helen N
中科院分区：
文献类型：
--
作者：
Troja, Weston;Kil, Kicheol;Klanke, Charles;Jones, Helen N
宫内生长受限（IUGR）是围产期并发症的主要原因，通常与胎盘脉管系统减少有关。最近的研究表明，IUGR 动物模型中 IGF-1 的过度表达可以维持胎盘脉管系统。然而，胎盘绒毛膜绒毛的细胞环境尚不清楚。体内滋养层和微血管内皮细胞的紧密接近暗示了 Ad-HuIGF-1 治疗后的自分泌/旁分泌调节。我们研究了 BeWo 绒毛膜癌和人胎盘微血管内皮细胞 (HPMVEC) 的共培养对内皮血管生成谱的影响，以及 Ad-HuIGF-1 处理一种细胞对另一种细胞的影响。 HPMVEC 从人类足月胎盘中分离出来，并在 37°C、5% CO2 的 EGM-2 中培养。 BeWo 细胞维持在 Ham 的 F12 营养混合物中，其中含有 10% FBS 和 1% pen/strep。将共培养的 HPMVECS+BeWo 细胞在无血清对照培养基、Ad-HuIGF-1 或 Ad-LacZ 中以 MOI 0 和 MOI 100:1 孵育 48 小时。将未经处理的细胞和单一培养的细胞与共培养的细胞进行比较。分别通过 RT-qPCR 和免疫细胞化学分析血管生成基因表达以及增殖和凋亡蛋白表达。使用学生 t 检验进行统计分析，P <0.05 被认为是显着的。直接 Ad-HuIGF-1 治疗增加 HPMVEC 增殖 (n =4) 并减少细胞凋亡 (n =3)。与单独培养的 HPMVEC 相比，共培养 HPMVEC+BeWo 细胞显着改变了血管生成谱的 RNA 表达 (n =8)。直接 Ad-HuIGF-1 处理显着增加 BeWo 细胞中的 Ang-1 (n =4)。 Ad-HuIGF-1 对 HPMVEC 的处理不会改变血管生成因子的 RNA 表达。滋养细胞因子可能在胎盘血管发育中起关键作用，IGF-1 可能在 HPMVEC 生长中起重要作用。胎盘绒毛膜绒毛的细胞环境相对未知。人胎盘微血管内皮细胞和模型滋养层细胞的共培养证明了两件事：微血管内皮细胞中HuIGF-1的过度表达改变了细胞生长周期，滋养层因子改变了微血管内皮细胞的血管生成特征。
Intrauterine growth restriction (IUGR) is a leading cause of perinatal complications, and is commonly associated with reduced placental vasculature. Recent studies demonstrated over‐expression of IGF‐1 in IUGR animal models maintains placental vasculature. However, the cellular environment of the placental chorionic villous is unknown. The close proximity of trophoblasts and microvascular endothelial cells in vivo alludes to autocrine/paracrine regulation following Ad‐HuIGF‐1 treatment. We investigated the co‐culturing of BeWo Choriocarcinoma and Human Placental Microvascular Endothelial Cells (HPMVECs) on the endothelial angiogenic profile and the effect Ad‐HuIGF‐1 treatment of one cell has on the other. HPMVECs were isolated from human term placentas and cultured in EGM‐2 at 37°C with 5% CO2. BeWo cells were maintained in Ham's F12 nutrient mix with 10% FBS and 1% pen/strep. Co‐cultured HPMVECS+BeWo cells were incubated in serum‐free control media, Ad‐HuIGF‐1, or Ad‐LacZ at MOI 0 and MOI 100:1 for 48 h. Non‐treated cells and mono‐cultured cells were compared to co‐cultured cells. Angiogenic gene expression and proliferative and apoptotic protein expression were analysed by RT‐qPCR and immunocytochemistry, respectively. Statistical analyses was performed using student's t‐test with P <0.05 considered significant. Direct Ad‐HuIGF‐1 treatment increased HPMVEC proliferation (n =4) and reduced apoptosis (n =3). Co‐culturing HPMVECs+BeWo cells significantly altered RNA expression of the angiogenic profile compared to mono‐cultured HPMVECs (n =8). Direct Ad‐HuIGF‐1 treatment significantly increased Ang‐1 (n =4) in BeWo cells. Ad‐HuIGF‐1 treatment of HPMVECs did not alter the RNA expression of angiogenic factors. Trophoblastic factors may play a key role in placental vascular development and IGF‐1 may have an important role in HPMVEC growth. Cellular environment of the placental chorionic villous is relatively unknown. Co‐culture of human placental microvascular endothelial cells and model trophoblasts demonstrated two things: the overexpression of HuIGF‐1 in the microvascular endothelial cells alters the cell growth cycle, and trophoblastic factors alter the angiogenic profile of the microvascular endothelial cells.