Abstract
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.
Highlights
In the human placenta following terminal villous differentiation, multiple cell types exist in close proximity (Gude et al 2004) and may undergo paracrine regulation following an insult or treatment of another cell type
This study investigated the effects of co-culturing BeWo cells and human placenta microvascular endothelial cells (HPMVECs) on angiogenic profile and if treatment of one cell type by adenoviral-mediated human Insulin-like Growth Factor-1 (HuIGF-1) impacted cellular responses of the other
Based on the percent of Ki67 expressed per field of view, direct Ad-HuIGF-1 treatment on Human Placental Microvascular Endothelial Cells (HPMVECs) significantly increased the proliferation of HPMVECs in mono-culture (P = 0.028, n = 4, Fig. 2B and C) compared to control untreated HPMVECs (Fig. 2A and C) suggesting IGF-1 may effect HPMVEC growth cycle
Summary
In the human placenta following terminal villous differentiation, multiple cell types exist in close proximity (Gude et al 2004) and may undergo paracrine regulation following an insult or treatment of another cell type. Previous studies have used conditioned medium to model paracrine signaling between multiple placental cell types associated with establishment of placentation and invasion on the maternal side, the models do not demonstrate the co-culture of two cell types in close proximity in vivo, nor do any represent the villous tree of the fetal side of the placenta (Zhou et al 2003). Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.
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