An improved purification approach with high cell viability and low cell loss for cryopreserved hepatocytes.
A modified purification procedure is described for effectively eliminating dead cells after hepatocyte cryopreservation. Isolated hepatocytes from six pig tissue samples were cryopreserved in liquid nitrogen for 2 weeks. After thawing, we developed a pre-incubation step prior to gradient centrifugation. The hepatocytes were subsequent cultured in suspension overnight (12-16 h), and then dead cells were eliminated by Ficoll 400 purification. The results showed that a high viability (mean of 96%) of cells was obtained, with a low viable cell loss in number (2-5%), by using this modified method.
Meng
Fan-ying
FY
Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. 963cat@sohu.com
Chen
Zhi-shui
ZS
Han
Meng
M
Hu
Xin-peng
XP
Zhou
Ping
P
D000818:Animals; D002452:Cell Count; D016923:Cell Death; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002499:Centrifugation, Density Gradient; D015925:Cryopreservation / Q000379:methods*; D005362:Ficoll; D022781:Hepatocytes / Q000166:cytology; D066298:In Vitro Techniques; D013552:Swine
D005362:Ficoll