A new method of combined cryopreservative hepatocytes / 临床外科杂志

ABSTRACT

Objective To find a preferable cryopreservative solution and enhance the viability further.Methods Donor hepatocytes were isolated from hybrid pig weighting 5～8 kg by means of collagenase.Then the hepatocyte suspension was put into a liquid nitrogen container for one month with three different cryoprotectants (only use DMSO,combined use of DMSO+HES and combined use of DMSO+HES+1.3 mmol/L Ca 2+).The samples were thawed rapidly and detected in the viability and morphology after a month.Fractional hepatocytes were cultured five days in containing Ca 2+ and uncontaining Ca 2+ culture media to observe the expression of function.Results Porcine hepatocytes,which were preserved in different culture media,show the different viability and morphology manifestation.Among the total the method of only uses DMSO show worst effect.Then is the DMSO+HES group.Combined cryopreservative method with HES+DMSO+1.3 mmol/LCacl 2 has a best effect and functional expression.Hepatocytes were cultured in culture media containing Ca 2+is better than those were cultured in culture media uncontaining Ca 2+.Conclusion The combined cryopreservative system in which HES+DMSO+1.3 mmol/LCacl 2 as the cardinal component can improve the preservative quality significantly.