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In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells
Published on: March 16, 2017
Bovine trophoblastic cell differentiation and binucleation involves enhanced endogenous retrovirus element expression
Katsuo Koshi1, Yasunori Suzuki, Yuki Nakaya
1Laboratory of Veterinary Physiology, Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan.
Bovine endogenous retrovirus (ERV) envelope genes, specifically bERVE-A and BERV-K1 env, are involved in bovine trophoblastic cell binucleation. Their expression increases during the formation of binucleate cells (BNCs) in vitro.
Area of Science:
- Reproductive biology
- Genetics
- Cell biology
Background:
- Endogenous retrovirus (ERV) envelope (env) genes play a role in trophoblast cell differentiation in humans and mice.
- Limited information exists regarding ERV roles in ruminant trophoblastic cells.
- This study investigates ERV elements in bovine trophoblastic cell binucleation.
Purpose of the Study:
- To explore the potential roles of ERV elements in bovine trophoblastic cell binucleation.
- To analyze the expression of specific ERV elements (bERVE-A, bERVE-B, BERV-K1 env, BERV-K2 env) in bovine trophoblastic cells.
- To assess the impact of in vitro culture conditions on ERV expression and binucleate cell (BNC) formation.
Main Methods:
- Collected blastocysts, elongated embryos, and endometrial/fetal membrane tissues from Japanese Black cows (days 17-37 gestation).
- Quantified gene expression of four ERV elements in tissues and cultured bovine trophoblastic (BT) cell lines using qRT-PCR.
- Induced BNC formation in BT cell lines using on-Matrigel and on-collagen gel cultures, examining gene/protein expression via qRT-PCR and immunocytochemistry.
Main Results:
- bERVE-A, bERVE-B, BERV-K1 env, and BERV-K2 env were expressed in BT cell lines; bERVE-A and BERV-K1 env were detected in peri-implantation trophoblastic tissues.
- On-Matrigel culture enhanced BNC-specific gene/molecule expression in BT cells.
- Expression of bERVE-A and BERV-K1 env increased during on-Matrigel culturing, correlating with BNC-specific gene expression (P < 0.01).
Conclusions:
- bERVE-A and BERV-K1 env are implicated in bovine trophoblastic cell binucleation and BNC-specific gene expression.
- Increased ERV expression during BNC formation suggests a role in the development of binucleate cells from mononucleate precursors.
- The on-Matrigel culture system provides a useful in vitro model for studying bovine trophoblast cell lineages.
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