title: Aggregate and Microcarrier Cultures of Human Pluripotent Stem Cells in Stirred-Suspension Systems.
abstract: Pluripotent stem cells can differentiate to any cell type and contribute to damaged tissue repair and organ function reconstitution. The scalable culture of pluripotent stem cells is essential to furthering the use of stem cell products in a wide gamut of applications such as screening of candidate drugs and cell replacement therapies. Human stem cell cultivation in stirred-suspension vessels enables the expansion of stem cells and the generation of differentiated progeny in quantities suitable for use in animal models and clinical studies. We describe methods of culturing human pluripotent stem cells in spinner flasks either as aggregates or on microcarriers. Techniques for assessing the quality of the culture and characterizing the cells based on the presentation of pertinent markers are also presented. Spinner flask culture with its relatively low capital and operating costs is appealing to laboratories interested in scaling up their production of stem/progenitor cells.
authors.affiliation: Department of Chemical and Biological Engineering, Tufts University, 4 Colby Street, 276A, Medford, MA, 02155, USA.
authors.forename: Preeti
authors.initials: P
authors.lastname: Ashok
authors.affiliation: Department of Chemical and Biological Engineering, Tufts University, 4 Colby Street, 276A, Medford, MA, 02155, USA.
authors.forename: Yongjia
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Department of Chemical and Biological Engineering, Tufts University, 4 Colby Street, 276A, Medford, MA, 02155, USA.
authors.forename: Mahboubeh R
authors.initials: MR
authors.lastname: Rostami
authors.affiliation: Department of Chemical and Biological Engineering, Tufts University, 4 Colby Street, 276A, Medford, MA, 02155, USA. Emmanuel.Tzanakakis@tufts.edu.|Tufts Clinical and Translational Science Institute, Tufts Medical Center, Boston, MA, 02111, USA. Emmanuel.Tzanakakis@tufts.edu.
authors.forename: Emmanuel S
authors.initials: ES
authors.lastname: Tzanakakis
mesh_terms: D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D004707:Endoderm / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007621:Karyotyping / Q000379:methods; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D012313:RNA
keywords: Cell aggregates; Differentiation; Microcarriers; Pluripotent stem cells; Scalable culture; Spinner flasks; Xeno-free culture