Growth of recombinant fibroblasts in alginate microcapsules.

摘要: To develop a novel strategy of nonautologous somatic gene therapy, we now demonstrate the feasibility culturing genetically modified fibroblasts within an immunoprotective environment and optimal conditions required for their continued survival in vitro. When mouse Ltk(-) transfected with human growth hormone were enclosed permselective microcapsules fabricated from alginate-polylysine-alginate, they to secrete at same rates as nonencapsulated cells. They also proliferate vitro least 1 month even though viability gradually declined about 50%. The can be improved by controlling (a) temperature during encapsulation, (b) duration treatment polylysine, (c) liquefying core alginate sodium citrate, (d) cell density time encapsulation. best leading maximum proliferation cells obtained encapsulating 4 10 degrees C instead room temperature, coating microspheres polylysine 6 min 20 min, treating citrate using concentration 2 x 10(6) cells/mL Under such conditions, normally adherent engineered survived proliferated optimally microcapsule environment. encapsulated maintained level transgene expression while recombinant products could diffuse through membrane without impediment. demonstration that survive continue deliver these optimization maximal should increase potential success microencapsulated therapy. 1994 John Wiley & Sons, Inc.