Senescent cultures of human dermal fibroblasts modified phenotype when immobilized in fibrin polymer.
One of the limitations in tissue engineering is the restricted ability to expand the number of cells, because somatic cells can duplicate a limited number of times before they lose the ability to divide, leading to a senescent state. Here we report that the interaction of senescent fibroblasts with fibrin polymer can modify the senescent phenotype and partially restore the ability of growth-arrested cells to continue replicating. Primary human dermal fibroblasts were grown to >90% SA/beta-Gal (senescence associated beta-galactosidase). The senescent cells were immobilized in fibrin-polymers by mixing fibrinogen and thrombin solutions. Immobilized senescent cell cultures grew, however, their growth arrested after 24 h of immobilization. The percentage of cells with a positive reaction at SA/beta-Gal did not decrease significantly after immobilization, but the intensity of the stain decreased. The glycolytic activity in immobilized senescent fibroblast was re-established at pre-senescent levels. In conclusion, fibrin induces changes in the phenotype of senescent human fibroblasts. This simple procedure could complement available tissue-engineering techniques to increase the amount of biomass seeded on a fibrin scaffold, which could be beyond senescence.
Universidad Técnica Federico Santa María, Biotechnology Center, Avenida España 1680, Valparaíso, Chile. cristian.acevedo@usm.cl
Cristian A
CA
Acevedo
Donald I
DI
Brown
Manuel E
ME
Young
Juan G
JG
Reyes
D002454:Cell Differentiation; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology*; D005337:Fibrin / Q000737:chemistry* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D006019:Glycolysis; D006801:Humans; D010641:Phenotype; D011108:Polymers / Q000737:chemistry*; D012867:Skin / Q000378:metabolism*
D011108:Polymers; D005337:Fibrin