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).

MATERIALI

Numero di prodotto

Marchio

Descrizione del prodotto

H3375

Sigma-Aldrich

HEPES, ≥99.5% (titration)

A6964

Sigma-Aldrich

Accutase^®, sterile-filtered, suitable for cell culture

A2153

Sigma-Aldrich

Sieroalbumina, lyophilized powder, ≥96% (agarose gel electrophoresis)

M3148

Sigma-Aldrich

2-mercaptoetanolo, Molecular Biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)

D9542

Sigma-Aldrich

DAPI, for nucleic acid staining

D5025

Sigma-Aldrich

Desossiribonucleasi I, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein

SML1046

Sigma-Aldrich

CHIR99021, ≥98% (HPLC)

P4543

Sigma-Aldrich

Valproic acid sodium salt, 98%

420099

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 (K_i= 5 nM) and Tyk2 (IC₅₀ = 1 nM).