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Macrophages modulate the growth and differentiation of rhesus monkey embryonic trophoblasts.

ABSTRACT: Immune cells within the endometrium at implantation are thought to play an important role in implantation, although their exact role is not well understood.A co-culture system of rhesus monkey embryos and maternal immune cells was established. Blastocysts obtained by in vitro fertilization were co-cultured with peripheral blood cells or decidual macrophages. Culture media were collected to assess secretions. Embryo growth was monitored, and trophoblasts were evaluated for proliferation, apoptosis, and differentiation.Embryonic trophoblast outgrowths were visible within 6 days of culture, and the area of embryo outgrowth was reduced when blastocysts were cultured with peripheral-derived or decidual macrophages. Trophoblast proliferation was not significantly affected with macrophage co-culture while chorionic gonadotropin secretion was increased. Trophoblast expression of CDH 11 and GJA1 was increased, suggesting that macrophages accelerate differentiation of peri-implantation trophoblasts.These results indicate an important role of macrophages in placentation and pregnancy success.

SUBMITTER: Rozner AE

PROVIDER: S-EPMC5056160 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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Macrophages modulate the growth and differentiation of rhesus monkey embryonic trophoblasts.

Rozner Ann E AE Durning Maureen M Kropp Jenna J Wiepz Gregory J GJ Golos Thaddeus G TG

American journal of reproductive immunology (New York, N.Y. : 1989) 20160917 5

<h4>Problem</h4>Immune cells within the endometrium at implantation are thought to play an important role in implantation, although their exact role is not well understood.<h4>Method of study</h4>A co-culture system of rhesus monkey embryos and maternal immune cells was established. Blastocysts obtained by in vitro fertilization were co-cultured with peripheral blood cells or decidual macrophages. Culture media were collected to assess secretions. Embryo growth was monitored, and trophoblasts we ...[more]

PMID: 27637575

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Project description:Embryonic stem cells (ESCs) may be able to cure or alleviate the symptoms of various degenerative diseases. However, unresolved issues regarding apoptosis, maintaining function and tumor formation mean a prudent approach should be taken towards advancing ESCs into human clinical trials. The rhesus monkey provides the ideal model organism for developing strategies to prevent immune rejection and test the feasibility, safety and efficacy of ESC-based medical treatments. Transcriptional profiling of rhesus ESCs provides a foundation for future pre-clinical ESC research using non-human primates as the model organism. In this research we use microarray, immunocytochemistry, real-time and standard RT-PCR to characterize and transcriptionally profile rhesus monkey embryonic stem cells. We identify 367 rhesus monkey stemness genes, we demonstrate the high level (>85%) of conservation of rhesus monkey stemness gene expression across five different rhesus monkey embryonic stem cell lines, we demonstrate that rhesus monkey ESC lines maintain a pluripotent undifferentiated state over a wide range of Pou5f1 (Oct-4) expression levels and we compare rhesus monkey, human and murine stemness genes to identify the key mammalian stemness genes. The supplementary tables list the genes that have been upregulated in each undifferentiated rhesus monkey embryonic stem cell line (GSM99998, GSM99999,GSM100000, GSM100001, GSM100002, GSM99965, GSM99966) in comparison analysis with the pooled differentiated embryonic stem cells (GSM99840). Supplemental Table 1 contains the comparison analysis for all 52,865 probe sets on the rhesus monkey gene chip, Supplemental Table 2 contains the rhesus monkey genes that were significantly upregulated (FC>3) in the ORMES-6 biological replicates, Supplemental Table 3 contains the rhesus monkey genes that were significantly upregulated (FC>3) in the pooled differentiated EBs and Supplemental Tables 4-8 represent genes that were significantly upregulated in ORMES 6A, 7, 9, 10 and 13 respectively. Supplemental Table 9 contains the RT-PCR primers used in this project. Keywords: Rhesus monkey embryonic stem cell microarray

Project description:Trophoblast differentiation during early placental development is critical for successful pregnancy and aberrant differentiation causes preeclampsia and early pregnancy loss. During the first trimester, cytotrophoblasts are exposed to low oxygen tension (equivalent to~2%-3% O2) and differentiation proceeds along an extravillous pathway (giving rise to invasive extravillous cytotrophoblasts) and a villous pathway (giving rise to multinucleated syncytiotrophoblast). Interstitial extravillous cytotrophoblasts invade the decidua, while endovascular extravillous cytotrophoblasts are involved in re-modelling uterine spiral arteries. We tested the idea that sodium butyrate (an epigenetic modulator) induces trophoblast differentiation in early gestation rhesus monkey trophoblasts through activation of the Wnt/β-catenin pathway. The results show that syncytiotrophoblast formation was increased by butyrate, accompanied by nuclear accumulation of β-catenin, and increased expression of EnvV2 and galectin-1 (two factors thought to be involved in trophoblast fusion). Surprisingly, the expression of GCM1 and syncytin-2 was not affected by sodium butyrate. When trophoblasts were incubated with lithium chloride, a GSK3 inhibitor that mimics Wnt activation, nuclear accumulation of β-catenin also occurred but differentiation into syncytiotrophoblast was not observed. Instead the cells differentiated to mononucleated spindle-shaped cells and showed molecular and behavioral characteristics of endovascular trophoblasts. Another highly specific inhibitor of GSK3, CHIR99021, failed to induce endovascular trophoblast characteristics. These observations suggest that activation of the Wnt/β-catenin pathway correlates with both trophoblast differentiation pathways, but that additional factors determine specific cell fate decisions. Other experiments suggested that the differential effects of sodium butyrate and lithium chloride might be explained by their effects on TNFα production. The results provide valuable tools to manipulate trophoblast differentiation in vitro and to better understand the differentiation pathways that occur during early gestation.

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Macrophages modulate the growth and differentiation of rhesus monkey embryonic trophoblasts.

Publications

Macrophages modulate the growth and differentiation of rhesus monkey embryonic trophoblasts.

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