Abstract
Rat hepatocytes were cryopreserved in hormonally-defined medium (HDM) containing either fetal bovine serum (FBS), glycerol, dimethyl sulfoxide (DMSO), sucrose or a mixture of these as a cryoprotectant. The best survival was with 10% (v/v) DMSO containing 30% (v/v) FBS using 5 × 105 hepatocytes ml−1 at −70 °C for 5 d on type I collagen-coated dishes. After thawing, the cell viability was 81% determined by the MTT-test. The cryopreserved hepatocytes had the capacity of albumin synthesis similar to hepatocytes without cryopreservation. This result shows that cryopreservation of rat hepatocyte can be used for the evaluation of hepatic functions.
Similar content being viewed by others
References
Alexandre E, Viollon-Abadie C, David P, Gandillet A, Coassolo P, Heyd B, Mantion G, Wolf P, Bachellier P, Jeack D, Richert L (2002) Cryopreservation of adult human hepatocytes obtained from resected liver biopsies. Cryobiology 44: 103–113.
Baust JM, Buskirk RV, Baust JG (2002) Modulation of the cryopreservation cap: elevated survival with reduced dimetyl sulfoxide concentration. Cryobiology 45: 97–108.
Darr TB, Hubel A (1997) Freezing characteristics of isolated pig and human hepatocytes. Cell Transplant. 6: 173–183.
Guillouzo A, Rialland L, Fautrel A, Guyomard C (1999) Survival and function of isolated hepatocytes after cryopreservation. Chem. Biol. Interact. 121: 7–16.
Hubálek Z (2003) Protectants used in the cryopreservation of microorganisms. Cryobiology 46: 205–229.
Kim SK, Yu SH, Lee JH, Lee JY, Rademacher A, Lee DH, Park JK (2001) Effect of collagen concentration on the viability and metabolic function of encapsulated hepatocytes. J. Microbiol. Biotechnol. 11: 423–427.
Lambert JJ, Coger RN (2001) Permeability of a model hepatocyte membrane to cryoprotectants at low temperatures. In: Proceedings of Bioengineering Conference ASME 2001, June 27-July 1, Utah, BED-Vol. 50, pp. 53–54.
Li AP (1999) Overview: hepatocytes and cryopreservation personal historical perspective. Chem. Biol. Interact. 121: 1–5.
Li AP, Jurima-Romet M (1997) Applications of primary human hepatocytes in the evaluation of pharmacokinetic drug-drug interactions: evaluation of model drugs terfenadine and rifampin. Cell Biol. Toxicol. 13: 365–374.
Merrill JC, Beck DJ, Kaminski DL, Li AP (1995) Polybrominated biphenyl induction of cytochrome P450 mixed function oxidase activity in primary rat and human hepatocytes. Toxicology 99: 147–152.
Seglen PO (1976) Preparation of rat liver cells. Meth. Biol. 13: 29–83.
Sugiyama N, Mizuguchi T, Aoki T, Hui T, Inderbitzin D, Demetriou AA, Rozga J (2002) Glycerol suppresses proliferation of rat hepatocytes and human HepG2 cells. J. Surg. Res. 103: 236–242.
Yu SH, Buchholz R, Kim SK (1999) Encapsulation of rat hepatocyte spheroids for the development of artificial liver. Biotechnol. Tech. 13: 609–614.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Son, J.H., Kim, KH., Nam, YK. et al. Optimization of cryoprotectants for cryopreservation of rat hepatocyte. Biotechnology Letters 26, 829–833 (2004). https://doi.org/10.1023/B:BILE.0000025886.57547.04
Issue Date:
DOI: https://doi.org/10.1023/B:BILE.0000025886.57547.04