Inoculation and growth conditions for high-cell-density expansion of mammalian neural stem cells in suspension bioreactors

Kallos, Michael S.; Behie, Leo A.

doi:10.1002/(sici)1097-0290(19990520)63:4<473::aid-bit11>3.0.co;2-c

Cited by 104 publications

(31 citation statements)

References 25 publications

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“…[33][34][35][36][37][38][39] Very low oxygen concentrations are often reported to foster greater retention of desirable stem cell behaviors, for instance, proliferation. However, culturing cells at low oxygen concentrations exacerbates the rapid depletion from flowing culture medium.…”

Section: Introductionmentioning

confidence: 99%

Transport and shear in a microfluidic membrane bilayer device for cell culture

2011

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Microfluidic devices have been established as useful platforms for cell culture for a broad range of applications, but challenges associated with controlling gradients of oxygen and other soluble factors and hemodynamic shear forces in small, confined channels have emerged. For instance, simple microfluidic constructs comprising a single cell culture compartment in a dynamic flow condition must handle tradeoffs between sustaining oxygen delivery and limiting hemodynamic shear forces imparted to the cells. These tradeoffs present significant difficulties in the culture of mesenchymal stem cells ͑MSCs͒, where shear is known to regulate signaling, proliferation, and expression. Several approaches designed to shield cells in microfluidic devices from excessive shear while maintaining sufficient oxygen concentrations and transport have been reported. Here we present the relationship between oxygen transport and shear in a "membrane bilayer" microfluidic device, in which soluble factors are delivered to a cell population by means of flow through a proximate channel separated from the culture channel by a membrane. We present an analytical model that describes the characteristics of this device and its ability to independently modulate oxygen delivery and hemodynamic shear imparted to the cultured cells. This bilayer configuration provides a more uniform oxygen concentration profile that is possible in a single-channel system, and it enables independent tuning of oxygen transport and shear parameters to meet requirements for MSCs and other cells known to be sensitive to hemodynamic shear stresses.

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Section: Introductionmentioning

confidence: 99%

Transport and shear in a microfluidic membrane bilayer device for cell culture

2011

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“…Within our own laboratory, work with mouse cell lines has established foundation protocols for human studies although the methods have not been directly transferable. Specifically, successful expansion of murine neural stem cells as aggregates in suspension bioreactors (Kallos & Behie 1999;Gilbertson et al, 2006) enabled us to develop protocols for human neural stem cells (Baghbaderani et al, 2010). Similarly, expanding murine ESCs as aggregates (Cormier et al, 2006;zur Nieden et al, 2007) allowed us to successfully expand pluripotent human ESCs as aggregates in suspension bioreactors (Krawetz et al, 2010 …”

Section: Mouse Embryonic Stem Cells As a Modelmentioning

confidence: 99%

“…The development of protocols for bioreactor expansion of stem cells has progressed rapidly in the past decade. Groups have successfully cultured hematopoietic stem cells, neural stem cells, human pancreatic progenitor cells and more recently both mouse and human embryonic stem cells in suspension (Zandstra et al, 1994;Kallos et al, 1999;Chawla et al, 2006;Cormier et al, 2006;Krawetz et al, 2009;Kehoe et al, 2010). Despite the advances in bioreactor design, there is still a need for optimization and standardization of suspension culture protocols to ensure reproducible and predictable cell populations for use in clinical applications.…”

Section: Introductionmentioning

confidence: 99%

Bioprocess Development for the Expansion of Embryonic Stem Cells

Hunt¹,

Alfred²,

Rancourt³

et al. 2011

Embryonic Stem Cells - Basic Biology to Bioengineering

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“…In spheroid culture, contacts and interactions among cells were substituted for the cell-surface contacts in monolayer cultures (Tolbert et al 1980;Goetghebeur et al 1991). Moreover, the spheroid formation can provide protection from the shear forces produced in suspension cultures in stirred bioreactors (Kallos and Behie 1999;Peshwa et al 1993).…”

Section: Introductionmentioning

confidence: 99%

Efficient formation of cell spheroids using polymer nanofibers

et al. 2011

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Spheroid culture has been used for suspension cultures of anchorage-dependent cells. In this study, we developed a new method for the suspension cultures of anchorage-dependent animal cells using polymer nanofibers. Poly(lactic-co-glycolic acid) nanofibers (785 nm in average fiber-diameter, 88 μm in average fiber-length) fabricated by the electrospinning method were added to each suspension culture of human embryonic kidney 293 cells and human dermal fibroblasts. As compared to no addition of nanofibers to the suspension cultures, nanofibers enhanced cell spheroid formation, thereby reducing cell death resulting from a lack of cell adhesion. Efficient formation of spheroids in the presence of polymer nanofibers may be useful for the suspension cultures of anchorage-dependent cells.

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Inoculation and growth conditions for high-cell-density expansion of mammalian neural stem cells in suspension bioreactors

Cited by 104 publications

References 25 publications

Transport and shear in a microfluidic membrane bilayer device for cell culture

Transport and shear in a microfluidic membrane bilayer device for cell culture

Bioprocess Development for the Expansion of Embryonic Stem Cells

Efficient formation of cell spheroids using polymer nanofibers

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