Comparative analysis of preparation of human hepatocytes by the ethylenediamine tetraacetic acid and collagenase technique.
OBJECTIVE
To compare the viability of human hepatocytes dissociated by the ethylenediaminetetraacetic acid and collagenase techniques.


METHODS
Hepatocytes were prepared by dissociation of liver fragments obtained from hepatectomies performed for therapeutic purposes at the Service of Digestive Tract Surgery, Federal University of Triângulo Mineiro.


RESULTS
During the first 4 days of the experiment, 70% of the cells presented birefringent membranes and were not stained with 2% erythrosine, and were therefore considered to be viable. During the first 3 days, hepatocyte viability was on average 71% in the EDTA group and 76% in the collagenase group, with no significant difference between groups. No significant difference was observed between groups at any time. The secretion of albumin by the cultured hepatocytes was preserved up to the seventh day. Mean albumin secretion during the first 3 days was 50 microg/ml in the two groups and a reduction of albumin production was observed from the fourth to the seventh day. Again, no significant difference was observed between groups at any time.


CONCLUSIONS
Cell viability and preservation of albumin secretion by hepatocytes are similar for the EDTA and collagenase techniques.
Crema
Eduardo
E
Experimental Laboratory Technique of Surgery, Department of Surgery, Federal University of Triângulo Mineiro, Brazil. cremauftm@mednet.com.br
Werneck
Ana Márcia
AM
Martins Júnior
Aiodair
A
Ribeiro
Lara Beatriz Prata
LB
Lima
Tatiana Silva de
TS
Silva
Alex Augusto
AA
D000418:Albumins / Q000187:drug effects* / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D017364:Collagenases / Q000494:pharmacology*; D004492:Edetic Acid / Q000494:pharmacology*; D006498:Hepatectomy; D022781:Hepatocytes / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D008099:Liver / Q000166:cytology*; D019164:Liver, Artificial; D009926:Organ Preservation; D010477:Perfusion
D000418:Albumins; D004492:Edetic Acid; D017364:Collagenases