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  • Optimized protocol for naive human pluripotent stem cell-derived trophoblast induction.

Optimized protocol for naive human pluripotent stem cell-derived trophoblast induction.

STAR protocols (2021-11-12)
Shingo Io, Yoshiki Iemura, Yasuhiro Takashima
ABSTRACT

Human trophoblasts arise from the morula as trophectoderm, which differentiates into cytotrophoblast, syncytiotrophoblast, and extravillous trophoblast after implantation. Here, we present a robust step-by-step protocol to induce trophectoderm (TE) from naive human pluripotent stem cells (PSCs) corresponding to pre-implantation epiblast. Our culture system (TE induction and ACE condition) mimics the entire trophoblast development including the molecular events. For complete details on the use and execution of this protocol, please refer to Io et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
2-Mercaptoethanol, Molecular Biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)
Sigma-Aldrich
CHIR99021, ≥98% (HPLC)
Millipore
JAK Inhibitor I, JAK Inhibitor I, CAS 457081-03-7, is a potent, reversible, cell-permeable, and ATP-competitive inhibitor of JAK 1 (IC₅₀ = 15 nM), JAK2 (IC₅₀ = 1 nM), JAK3 (Ki = 5 nM) and Tyk2 (IC₅₀ = 1 nM).
Sigma-Aldrich
Accutase® solution, sterile-filtered, suitable for cell culture
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Valproic acid sodium salt, 98%
Sigma-Aldrich
Deoxyribonuclease I from bovine pancreas, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein