Human Pluripotent Stem Cell Expansion in Stirred Tank Bioreactors.

Automation Human pluripotent stem cells (hPSCs) Perfusion feeding Process development Stirred tank bioreactors (STBRs) Suspension culture

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2019
Historique:
entrez: 25 5 2019
pubmed: 28 5 2019
medline: 13 3 2020
Statut: ppublish

Résumé

This chapter describes a detailed protocol on human pluripotent stem cells (hPSCs) cultivation as matrix-free cell-only aggregates in defined and xeno-free culture medium in stirred tank bioreactors (STBRs). Starting with a frozen stock pre-expanded on conventional culture dishes (2D), the ultimate process is performed in 150 mL culture scale in stirred tank bioreactors (3D) and is designed to produce up to 500 million pluripotent hPSC within 7 days. The culture strategy includes perfusion-based cell feeding facilitating process control, automation, and higher cell yields. Ultimately, this detailed protocol describes an important step for generating a defined starting cell population for directed lineage differentiation and subsequently fueling human cell-based assays and regenerative medicine approaches.

Identifiants

pubmed: 31124106
doi: 10.1007/978-1-4939-9477-9_7
doi:

Substances chimiques

Culture Media 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

79-91

Auteurs

Felix Manstein (F)

Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiac, Thoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany.
REBIRTH-Cluster of Excellence, Hannover Medical School, Hannover, Germany.

Caroline Halloin (C)

Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiac, Thoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany.
REBIRTH-Cluster of Excellence, Hannover Medical School, Hannover, Germany.

Robert Zweigerdt (R)

Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiac, Thoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany. zweigerdt.robert@mh-hannover.de.
REBIRTH-Cluster of Excellence, Hannover Medical School, Hannover, Germany. zweigerdt.robert@mh-hannover.de.

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Classifications MeSH