Please use this identifier to cite or link to this item: https://doi.org/10.1002/jbm.a.31864
Title: Functional and phenotypic characterization of human keratinocytes expanded in microcarrier culture
Authors: Borg, D.J.
Dawson, R.A.
Leavesley, D.I.
Hutmacher, D.W. 
Upton, Z.
Malda, J.
Keywords: 3D culture
Human skin equivalent model
Keratinocyte
Microcarrier
Skin tissue engineering
Issue Date: Jan-2009
Source: Borg, D.J., Dawson, R.A., Leavesley, D.I., Hutmacher, D.W., Upton, Z., Malda, J. (2009-01). Functional and phenotypic characterization of human keratinocytes expanded in microcarrier culture. Journal of Biomedical Materials Research - Part A 88 (1) : 184-194. ScholarBank@NUS Repository. https://doi.org/10.1002/jbm.a.31864
Abstract: Skin cells for transplantation are routinely prepared by growing patient keratinocytes in a semi-defined cocktail of growth factors, including serum and feeder cells. However, these reagents require substantial risk remediation and can contribute to transplant rejection. Microcarrier culture is an emerging technology that may allow the elimination of feeder cells whilst facilitating expansion of cultured keratinocytes. However, the behavior of keratinocytes in microcarrier culture and the potential of these cells to form an epidermis have been poorly defined. We characterized freshly isolated human keratinocytes cultured on CultiSpher-G® microcarriers in the absence of murine feeder cells and assessed the potential of the keratinocytes to form an epidermis in an in vitro model. In a single passage, keratinocytes multiplied 44.9-fold in microcarrier-bioreactor culture in 17 days, whereas two-dimensional cultures reached confluence in 9 days and only expanded 7.4-fold. Histological characterization of keratinocytes on the microcarriers revealed that the cells were randomly distributed within these porous structures, however, not all pores contained cells. High-resolution microcomputed tomography imaging of the microcarriers confirmed limited intercorrnectivity of the pores. Immunoreactivity of specific epidermal markers was confirmed during cell expansion via immunohistochemistry. Despite the expression of differentiation markers, microcarrier-expanded keratinocytes retained the capacity to form an epidermis, as was evaluated using an in vitro human skin equivalent model. The epidermis formed by microcarrier-expanded keratinocytes in this model exhibited morphology similar to native skin. Significantly, the microcarrier technique successfully eliminates the need for a feeder cell layer and hence facilitates development of an improved culture system. © 2008 Wiley Periodicals, Inc.
Source Title: Journal of Biomedical Materials Research - Part A
URI: http://scholarbank.nus.edu.sg/handle/10635/67071
ISSN: 15493296
DOI: 10.1002/jbm.a.31864
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

22
checked on Dec 6, 2017

WEB OF SCIENCETM
Citations

19
checked on Nov 22, 2017

Page view(s)

27
checked on Dec 10, 2017

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.