Microencapsulation of CHO cells in a hydroxyethyl methacrylate-methyl methacrylate copolymer.
Chinese hamster ovary fibroblasts, as model cells, have been microencapsulated in a hydroxyethyl methacrylate-methyl methacrylate copolymer (HEMA-MMA) by interfacial precipitation. The polymer containing approximately equal to 75 mol% HEMA, dissolved in polyethylene glycol 200 (PEG 200) was coextruded with the cell suspension (4-6 X 10(5) cells/ml in the alpha-MEM with 10% foetal calf serum +/- Ficoll 400/PBS) through a concentric needle assembly. Polymer solution droplets, containing cells, were blown off the end of the needle assembly by a coaxial filtered air stream into a nonsolvent bath containing phosphate buffered saline (PBS) with 5 ppm Pluronic L101, overlaid with hexadecane. The nascent capsules hang at the hexadecane/PBS interface while the solvent is extracted into the aqueous nonsolvent, to precipitate the polymer around the cells. The resultant capsules were 500 microns-1 mm in diam. with a microporous sponge-like interior, and also very tough and flexible. The cells survived encapsulation based on subculture ability, retention of some fluorescein diacetate (FDA) activity over 5 d and direct light microscopic evidence of cell growth over 10 d after histological sectioning and staining. However, cell growth was not uniformly observed (especially in the FDA assay) and this was attributed to space limitations for growth within the microporous interior. Continued development of this process and adaptation to cells such as pancreatic islets is expected to lead to hybrid artificial organs which are capable of ameliorating metabolic disorders such as diabetes.
Dawson
R M
RM
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Ontario, Canada.
Broughton
R L
RL
Stevenson
W T
WT
Sefton
M V
MV
D000179:Acrylates; D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D004339:Drug Compounding; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D005452:Fluoresceins; D008689:Methacrylates; D020366:Methylmethacrylate; D008768:Methylmethacrylates; D010053:Ovary / Q000166:cytology*; D011108:Polymers
D000179:Acrylates; D005452:Fluoresceins; D008689:Methacrylates; D008768:Methylmethacrylates; D011108:Polymers; D020366:Methylmethacrylate; C005044:hydroxyethyl methacrylate; C018506:diacetylfluorescein