[The protective effect of Astragalus membranaceus extraction on cryopreserved primary-cultured human fetal hepatocytes].
OBJECTIVE
To observe the influence of Astragalus membranaceus Extraction (AE) on the primary-cultured human fetal hepatocytes stored in liquid nitrogen and explore a new method for the cryopreservation of human hepatocytes with improved function.


METHODS
Human fetal hepatocytes were harvested by two-step collagenase perfusion, and then stored in a liquid nitrogen for one month with five different cryoprotectants (I: 10% DMSO, II: 5% DMSO + 2 mg/L AE, III: 5% DMSO + 20 mg/L AE, IV: 5% DMSO + 60 mg/L AE, V: 5% DMSO + 100 mg/L AE). One month later, the cells were thawed rapidly and the viability, morphology and basic function of them were tested.


RESULTS
The human fetal hepatocytes in different groups showed various levels of viability, morphological manifestation and cell function respectively. After thawing, the viability rate and flash adhering rate in group IV and V had no significant difference with group I (P > 0.05), but were higher than group II and III (P < 0.05); the cell function analysis in the group IV, the results of ALB and AST level determination, NH4Cl transformation test, were the best among the groups (P < 0.05).


CONCLUSIONS
AE can provide protection for human fetal hepatocytes in cryopreservation, and the best performance concentration level of its is 60 mg/L; the preservation dosage of DMSO can be reduced when combined with AE in the preservation solution, which shows that AE has a synergistic effect with DMSO.
Wang
Yan
Y
The Southern Medical University Affiliated Zhujiang Hospital, Guangzhou 510282, China.
Zhu
Ying-qian
YQ
Lin
Jing-ming
JM
Zhang
Zhong-yi
ZY
Gao
Yi
Y
D000418:Albumins / Q000378:metabolism; D001219:Aspartate Aminotransferases / Q000378:metabolism; D027885:Astragalus propinquus / Q000737:chemistry; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D005333:Fetus; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D006801:Humans
D000418:Albumins; D003451:Cryoprotective Agents; D004365:Drugs, Chinese Herbal; D001219:Aspartate Aminotransferases; D004121:Dimethyl Sulfoxide