A method for preparation of hydrogel microcapsules for stem cell bioprocessing and stem cell therapy.
A method for the preparation of suspension culture microcapsules used in the bioprocessing of human mesenchymal stem cells (hMSCs) is reported. The microcapsules are prepared from a semi-synthetic hydrogel comprising Pluronic®F127 conjugated to denatured fibrinogen. The Pluronic-fibrinogen adducts display a lower critical solubility temperature (LCST) at ∼30 °C, thus enabling mild, cell-compatible physical crosslinking of the microcapsules in a warm gelation bath. Cell-laden microgels were prepared from a solution of Pluronic-fibrinogen hydrogel precursor and hMSCs; these were cultivated for up to 15 days in laboratory-scale suspension bioreactors and harvested by reducing the temperature of the microcapsules to disassemble the physical polymer network. The viability, proliferation and cell recovery yields of the hMSCs were shown to be better than photo-chemically crosslinked microcapsules made from a similar material. The cell culture yields, which exceeded 300% after 15 days in suspension culture, were comparable to other microcarrier systems used for the mass production of hMSCs. The simplicity of this methodology, both in terms of the cell inoculation and mild recovery conditions, represent distinct advantages for stem cell bioprocessing with suspension culture bioreactors.
The Faculty of Biomedical Engineering, Technion - Israel Institute of Technology, Haifa 32000, Israel; The Interdisciplinary Program for Biotechnology, Technion - Israel Institute of Technology, Haifa 32000, Israel.
Revital
R
Goldshmid
The Faculty of Biomedical Engineering, Technion - Israel Institute of Technology, Haifa 32000, Israel.
Iris
I
Mironi-Harpaz
The Faculty of Biomedical Engineering, Technion - Israel Institute of Technology, Haifa 32000, Israel.
Yonatan
Y
Shachaf
The Faculty of Biomedical Engineering, Technion - Israel Institute of Technology, Haifa 32000, Israel. Electronic address: dror@bm.technion.ac.il.
Dror
D
Seliktar
D019149:Bioreactors; D002214:Capsules / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D002470:Cell Survival; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D003432:Cross-Linking Reagents; D005340:Fibrinogen / Q000737:chemistry; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D020442:Poloxamer / Q000737:chemistry
D002214:Capsules; D003432:Cross-Linking Reagents; D020100:Hydrogels; D020442:Poloxamer; D005340:Fibrinogen
3D culture; Biomaterial; Chemical crosslinking; PEG; Physical crosslinking; Pluronic-fibrinogen; Suspension bioreactor