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  • A protocol to induce expandable limb-bud mesenchymal cells from human pluripotent stem cells.

A protocol to induce expandable limb-bud mesenchymal cells from human pluripotent stem cells.

STAR protocols (2022-11-02)
Tomoka Takao, Daisuke Yamada, Takeshi Takarada
ABSTRACT

Here, we present a protocol for the selective differentiation of human pluripotent stem cells mimicking human developmental processes into expandable PRRX1+ limb-bud mesenchymal (ExpLBM) cells. This approach enables expansion through serial passage while maintaining capacity for chondrogenic differentiation. For complete details on the use and execution of this protocol, please refer to Yamada et al. (2021, 2022).

MATERIALS
Product Number
Brand
Product Description

Roche
Insulin, human, recombinant (yeast)
Sigma-Aldrich
Anti-Sox9 Antibody, Chemicon®, from rabbit
Sigma-Aldrich
Human Plasma Fibronectin Purified Protein, from human plasma, liquid, 1 mg/mL (100 MG pack size is lyophilized), purified protein, suitable for cell culture
Sigma-Aldrich
PI 3-K inhibitor IX, PIK-90, The PI 3-K inhibitor IX, PIK-90, also referenced under CAS 677338-12-4, controls the biological activity of PI 3-K. This small molecule/inhibitor is primarily used for Phosphorylation & Dephosphorylation applications.