TY - JOUR
T1 - Stirred bioreactors for the expansion of adult pancreatic stem cells
AU - Serra, Margarida
AU - Brito, Catarina
AU - Leite, Sofia Batista
AU - Gorjup, Erwin
AU - von Briesen, Hagen
AU - Carrondo, Manuel J. T.
AU - Alves, Paula M.
N1 - Funding Information:
The authors acknowledge António Roldão and Marcos Sousa for helpful discussions on this manuscript. The work was supported by the European Commission (Cell Programming by Nanoscaled Devices- NMP4-CT-2004-500039).
Copyright:
Copyright 2009 Elsevier B.V., All rights reserved.
PY - 2009/1
Y1 - 2009/1
N2 - Adult pluripotent stem cells are a cellular resource representing unprecedented potential for cell therapy and tissue engineering. Complementary to this promise, there is a need for efficient bioprocesses for their large scale expansion and/or differentiation. With this goal in mind, our work focused on the development of three-dimensional (3-D) culture systems for controlled expansion of adult pancreatic stem cells (PSCs). For this purpose, two different culturing strategies were evaluated, using spinner vessels: cell aggregated cultures versus microcarrier technology. The use of microcarrier supports (Cytodex 1 and Cytodex 3) rendered expanded cell populations which retained their self-renewal ability, cell marker, and the potential to differentiate into adipocytes. This strategy surmounted the drawbacks of aggregates in culture which were demonstrably unfeasible as cells clumped together did not proliferate and lost PSC marker expression. Furthermore, the results obtained showed that although both microcarriers tested here were suitable for sustaining cell expansion, Cytodex 3 provided a better substrate for the promotion of cell adherence and growth. For the latter approach, the potential of bioreactor technology was combined with the efficient Cytodex 3 strategy under controlled environmental conditions (pH-7.2, pO2-30% and temperature-37 °C); cell growth was more efficient, as shown by faster doubling time, higher growth rate and higher fold increase in cell concentration, when compared to spinner cultures. This study describes a robust bioprocess for the controlled expansion of adult PSC, representing an efficient starting point for the development of novel technologies for cell therapy.
AB - Adult pluripotent stem cells are a cellular resource representing unprecedented potential for cell therapy and tissue engineering. Complementary to this promise, there is a need for efficient bioprocesses for their large scale expansion and/or differentiation. With this goal in mind, our work focused on the development of three-dimensional (3-D) culture systems for controlled expansion of adult pancreatic stem cells (PSCs). For this purpose, two different culturing strategies were evaluated, using spinner vessels: cell aggregated cultures versus microcarrier technology. The use of microcarrier supports (Cytodex 1 and Cytodex 3) rendered expanded cell populations which retained their self-renewal ability, cell marker, and the potential to differentiate into adipocytes. This strategy surmounted the drawbacks of aggregates in culture which were demonstrably unfeasible as cells clumped together did not proliferate and lost PSC marker expression. Furthermore, the results obtained showed that although both microcarriers tested here were suitable for sustaining cell expansion, Cytodex 3 provided a better substrate for the promotion of cell adherence and growth. For the latter approach, the potential of bioreactor technology was combined with the efficient Cytodex 3 strategy under controlled environmental conditions (pH-7.2, pO2-30% and temperature-37 °C); cell growth was more efficient, as shown by faster doubling time, higher growth rate and higher fold increase in cell concentration, when compared to spinner cultures. This study describes a robust bioprocess for the controlled expansion of adult PSC, representing an efficient starting point for the development of novel technologies for cell therapy.
KW - Adult stem cells
KW - Expansion
KW - Microcarriers
KW - Pancreatic stem cells
KW - Stirred bioreactors
UR - http://www.scopus.com/inward/record.url?scp=58149314043&partnerID=8YFLogxK
U2 - 10.1016/j.aanat.2008.09.005
DO - 10.1016/j.aanat.2008.09.005
M3 - Article
C2 - 19054658
SN - 0940-9602
VL - 191
SP - 104
EP - 115
JO - Annals Of Anatomy-Anatomischer Anzeiger
JF - Annals Of Anatomy-Anatomischer Anzeiger
IS - 1
ER -