Kiwifruit actinidin: a proper new collagenase for isolation of cells from different tissues.
Actinidin is a cysteine protease abundant in Kiwifruit. This enzyme is known as a meat-tenderizing protease. In this project, actinidin was purified from kiwifruit by salt precipitation and ion exchange chromatography. Collagenolytic effect of the purified enzyme was tested in four different buffer systems. Thereafter, the enzyme was used for isolation and culture of cells from three different tissues: endothelial cells from human umbilical vein, hepatocytes from rat liver, and thymic epithelial cells from rat thymus. Our results revealed that actinidin can hydrolyze collagen types I and II at neutral and alkaline buffers. Furthermore, actinidin compared with type II or IV collagenase isolated intact human umbilical vein endothelial cells, hepatocytes, and thymic epithelial cells with viability more than 90%. These results address a novel and valuable collagenase, which can be used efficiently for hydrolysis of collagen and isolation of different cell populations from various solid tissues.
Mostafaie
Ali
A
Medical Biology Research Center, Kermanshah University of Medical Sciences, Sorkheh Ligeh, P.O. Box 1568, Kermanshah, Iran. amostafaie@kums.ac.ir
Bidmeshkipour
Ali
A
Shirvani
Zeinab
Z
Mansouri
Kamran
K
Chalabi
Maryam
M
D029042:Actinidia / Q000201:enzymology*; D000818:Animals; D002469:Cell Separation; D002470:Cell Survival; D002478:Cells, Cultured; D017364:Collagenases / Q000737:chemistry* / Q000302:isolation & purification; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells / Q000166:cytology; D005638:Fruit / Q000201:enzymology*; D006801:Humans; D008099:Liver / Q000166:cytology / Q000187:drug effects; D051381:Rats; D013950:Thymus Gland / Q000166:cytology / Q000187:drug effects
D003546:Cysteine Endopeptidases; C018745:actinidain; D017364:Collagenases