Cryopreservation of fibroblasts immobilized within a porous scaffold: effects of preculture and collagen coating of scaffold on performance of three-dimensional cryopreservation.
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.
Miyoshi
Hirotoshi
H
Department of Biomedical Engineering, Institute of Basic Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan. hmiyoshi@md.tsukuba.ac.jp
Ehashi
Tomo
T
Ohshima
Norio
N
Jagawa
Akari
A
D000818:Animals; D049109:Cell Proliferation; D003094:Collagen / Q000737:chemistry; D015925:Cryopreservation / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology*; D051379:Mice; D041681:NIH 3T3 Cells; D016062:Porosity; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry
D003094:Collagen