title: Expansion of Human Mesenchymal Stem Cells in a Microcarrier Bioreactor.
abstract: Human mesenchymal stem cells (hMSCs) are considered as a primary candidate in cell therapy owing to their self-renewability, high differentiation capabilities, and secretions of trophic factors. In clinical application, a large quantity of therapeutically competent hMSCs is required that cannot be produced in conventional petri dish culture. Bioreactors are scalable and have the capacity to meet the production demand. Microcarrier suspension culture in stirred-tank bioreactors is the most widely used method to expand anchorage dependent cells in a large scale. Stirred-tank bioreactors have the potential to scale up and microcarriers provide the high surface-volume ratio. As a result, a spinner flask bioreactor with microcarriers has been commonly used in large scale expansion of adherent cells. This chapter describes a detailed culture protocol for hMSC expansion in a 125 mL spinner flask using microcarriers, Cytodex I, and a procedure for cell seeding, expansion, metabolic sampling, and quantification and visualization using microculture tetrazolium (MTT) reagent.
authors.affiliation: Department of Chemical and Biomedical Engineering, Florida State University, 2525 Pottsdamer St., Tallahassee, FL, 32310, USA.
authors.forename: Ang-Chen
authors.initials: AC
authors.lastname: Tsai
authors.affiliation: Department of Chemical and Biomedical Engineering, Florida State University, 2525 Pottsdamer St., Tallahassee, FL, 32310, USA. teng@eng.fsu.edu.
authors.forename: Teng
authors.initials: T
authors.lastname: Ma
mesh_terms: D019149:Bioreactors; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D004867:Equipment Design; D005947:Glucose / Q000032:analysis / Q000378:metabolism; D006801:Humans; D019344:Lactic Acid / Q000032:analysis / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D019344:Lactic Acid; D005947:Glucose
keywords: Bioreactor; Cell culture; Cell expansion; MSC; Mesenchymal stem cell; Microcarrier; Spinner flask