Nucleofection (Amaxa) and electroporation (Biorad) of hPSCs

This collection describes the standard procedure for the delivery of plasmids, mRNA or ribonucleoprotein (RNP) into human pluripotent stem cells (hPSCs) using nucleofection or electroporation.

Collection Overview

Preparing MEF-cultured hPSCs for nucleofection

Preparing Feeder-free hPSCs for nucleofection

Preparing plasmids for nucleofection

Preparing mRNA for nucleofection

In vitro assembling of RNP for nucleofection

Nucleofection of hPSCs

Electroporation of hPSCs

A. Preparation of MEFs-cultured hPSCs for electroporation

B. Preparing plasmids for electroporation

C. Electroporation

FACS-based enrichment of transfected cells

A. Preparation of samples for FACS sorting

B. After FACS

General notes

1. Throughout these protocols, the term hPSC is used to collectively refer to both hiPSCs and hESCs. All described procedures have been tested and work equally well for hiPSCs and hESCs.