Cryopreservation of Fibroblasts Immobilized Within a Porous Scaffold: Effects of Preculture and Collagen Coating of Scaffold on Performance of Three‐Dimensional Cryopreservation

Abstract

As a preliminary investigation to establish a cryopreservation method suited for bioartificial livers (BALs), three‐dimensional (3‐D) cryopreservation experiments with fibroblasts were performed, in which the cells were firstly seeded into a porous scaffold, and the scaffold containing the cells was then cryopreserved. After thawing, 65% of the initially applied cells were still attached to the scaffold, and this efficiency was significantly higher than that in the control experiments (39%), in which fibroblasts cryopreserved in a suspension were seeded into the scaffold. This higher efficiency was mainly caused by higher immobilization efficiency at the time of cell seeding (83%) than in the controls (54%). Collagen coating of the scaffold in the 3‐D cryopreservation enhanced immobilization efficiency at the time of cell seeding, and 1‐day precultures before the 3‐D cryopreservation considerably improved cell growth after thawing. From these favorable results, this 3‐D cryopreservation method may become useful for developing BALs.