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Merck
CN
  • 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
摘要

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).

材料
Product Number
品牌
产品描述

Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
Accutase® 溶液, sterile-filtered, suitable for cell culture
Sigma-Aldrich
2-巯基乙醇, Molecular Biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
脱氧核糖核酸酶 I 来源于牛胰腺, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein
Sigma-Aldrich
CHIR99021, ≥98% (HPLC)
Sigma-Aldrich
丙戊酸 钠盐, 98%
Millipore
JAK抑制剂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).