Expansion of mesenchymal stem cells using a microcarrier-based cultivation system: growth and metabolism.
For the continuous and fast expansion of mesenchymal stem cells (MSCs), microcarriers have gained increasing interest. The aim of this study was to evaluate the growth and metabolism profiles of MSCs, expanded in a microcarrier-based cultivation system. We investigated various cultivation conditions to expand goat mesenchymal stem cells on Cytodex 1 microcarriers. These conditions differed in feeding regime, i.e. the addition of fresh proliferation medium, with or without new microcarriers. For all conditions, cell attachment, cell proliferation, energy source consumption, metabolite production, and cell distribution on the microcarriers were studied. Attachment efficiencies of 40% were obtained followed by successful expansion up to 15 cultivation days. Depending on the feeding regime, an exponential growth, stationary growth, and decline growth phase could be distinguished. Addition of 30% fresh medium containing microcarriers every three days showed the longest continuous proliferation of goat MSCs on microcarriers. This feeding regime has the advantage that metabolites, such as ammonia, are diluted and that new energy sources, such as glucose and glutamine, and additional surface area are provided to the cells. In addition, by adding extra microcarriers a more homogenous cell distribution on the microcarriers is obtained as a result of bead-to-bead transfer. A correlation between nutrient consumption, metabolite production and cell growth was observed. The decreasing yield of lactate from glucose over time indicated a possible shift in cellular metabolism.
Schop
D
D
Xpand Biotechnology BV, Bilthoven, The Netherlands. Deborah.Schop@progentix.com
Janssen
F W
FW
Borgart
E
E
de Bruijn
J D
JD
van Dijkhuizen-Radersma
R
R
D000641:Ammonia / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects; D003911:Dextrans / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D006041:Goats; D019344:Lactic Acid / Q000096:biosynthesis; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors
D003911:Dextrans; D005973:Glutamine; D019344:Lactic Acid; D000641:Ammonia; C052839:Cytodex; D005947:Glucose