Mass cultivation of human retinal pigment epithelial cells with microcarrier.
Microcarrier cell culture permits mass cultivation of anchorage-dependent cells. In this study, mass cultivation of human retinal pigment epithelial (RPE) cells was studied using Cytodex 3 (Pharmacia) as a microcarrier. Human RPE cells were established from aborted fetuses and cultured in Dulbecco's modified Eagle's Medium (DMEM). After the 3rd-5th passages, RPE cells were suspended in 50 ml of DMEM in a spinner flask at a density of 2 x 10(5)/ml, and Cytodex 3 was added to the spinner flask at a bead density of 10 mg/ml. Cultures were maintained at 20-50 rpm (final speed) on a magnetic stirrer, and DMEM was added up to 100 ml. Fifty milliliters of DMEM were decanted and replaced with fresh DMEM every 2 days. After 1 week, a cell density of 10(6)/ml DMEM was obtained. Phase contrast microscopy showed bridging formation between microcarriers, which suggests tight cell adhesion. Microcarrier cell culture has a variety of advantages which include greater cell production, use of less medium and less risk of contamination compared to the conventional monolayer culture technique, and it also allows passaging without using proteases. Using this culture system, greater possibilities for wider application of new cell cultures can be expected.
D002452:Cell Count; D002455:Cell Division; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D003911:Dextrans; D005333:Fetus; D006801:Humans; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000648:ultrastructure
D003911:Dextrans; C052839:Cytodex