title: Compartmental hollow fiber capillary membrane-based bioreactor technology for in vitro studies on red blood cell lineage direction of hematopoietic stem cells.
abstract: Continuous production of red blood cells (RBCs) in an automated closed culture system using hematopoietic stem cell (HSC) progenitor cell populations is of interest for clinical application because of the high demand for blood transfusions. Previously, we introduced a four-compartment bioreactor that consisted of two bundles of hollow fiber microfiltration membranes for transport of culture medium (forming two medium compartments), interwoven with one bundle of hollow fiber membranes for transport of oxygen (O(2)), carbon dioxide (CO(2)), and other gases (forming one gas compartment). Small-scale prototypes were developed of the three-dimensional (3D) perfusion cell culture systems, which enable convection-based mass transfer and integral oxygenation in the cell compartment. CD34(+) HSC were isolated from human cord blood units using a magnetic separation procedure. Cells were inoculated into 2- or 8-mL scaled-down versions of the previously designed 800-mL cell compartment devices and perfused with erythrocyte proliferation and differentiation medium. First, using the small-scale 2-mL analytical scale bioreactor, with an initial seeding density of 800,000 cells/mL, we demonstrated approximately 100-fold cell expansion and differentiation after 7 days of culture. An 8-mL laboratory-scale bioreactor was then used to show pseudocontinuous production by intermediately harvesting cells. Subsequently, we were able to use a model to demonstrate semicontinuous production with up to 14,288-fold expansion using seeding densities of 800,000 cells/mL. The down-scaled culture technology allows for expansion of CD34(+) cells and stimulating these progenitors towards RBC lineage, expressing approximately 40% CD235(+) and enucleation. The 3D perfusion technology provides an innovative tool for studies on RBC production, which is scalable.
authors.affiliation: Department of Surgery, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15203, USA.
authors.forename: Greggory J
authors.initials: GJ
authors.lastname: Housler
authors.forename: Toshio
authors.initials: T
authors.lastname: Miki
authors.forename: Eva
authors.initials: E
authors.lastname: Schmelzer
authors.forename: Christopher
authors.initials: C
authors.lastname: Pekor
authors.forename: Xiaokui
authors.initials: X
authors.lastname: Zhang
authors.forename: Lin
authors.initials: L
authors.lastname: Kang
authors.forename: Vanessa
authors.initials: V
authors.lastname: Voskinarian-Berse
authors.forename: Stewart
authors.initials: S
authors.lastname: Abbot
authors.forename: Katrin
authors.initials: K
authors.lastname: Zeilinger
authors.forename: Jörg C
authors.initials: JC
authors.lastname: Gerlach
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D000954:Antigens, Surface / Q000378:metabolism; D019149:Bioreactors; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002454:Cell Differentiation; D019070:Cell Lineage; D002470:Cell Survival; D002478:Cells, Cultured; D004912:Erythrocytes / Q000166:cytology* / Q000648:ultrastructure; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005947:Glucose / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D019344:Lactic Acid / Q000096:biosynthesis; D008567:Membranes, Artificial; D010477:Perfusion; D013194:Staining and Labeling; D013997:Time Factors
chemical_list: D000180:Acrylic Resins; D000954:Antigens, Surface; D008567:Membranes, Artificial; D019344:Lactic Acid; D005947:Glucose