Work

Nucleofection (Amaxa) and electroporation (Biorad) of hPSCs v2

Year: 2023

Type: preprint

Abstract: 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 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.

DOI: 10.17504/protocols.io.e6nvwkkewvmk/v2

Authors: Hanqin Li, Oriol Busquets, Steven Poser, Dirk Hockemeyer, Frank Soldner

Institutions: Albert Einstein College of Medicine, University of California, Berkeley

Language: English

Topic: Pluripotent Stem Cells Research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: green

OpenAlex ID: W4315783129

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Cited by: 1

Related to: 10