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KMID : 0376119940210010075
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Medical Journal of the Red Cross Hospital
1994 Volume.21 No. 1 p.75 ~ p.90
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The Isolation of Rabbit Glomeruli by Iron Oxide Particle Perfusion Method and Mesangial Cell Culture
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Abstract
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It is well known that kidney mesangial cells have many functions. Such as maintenance of glomerular capillary microcirculation, regulation of GFR, production of various growith factors and cytodines, and formation of collagen and extracellular
matrix.
So it is regarded that mesangial cells will be involved and play important roles in the pathogenesis of many glomerular diseases.
Mesangial cell cultrue is thought to be one of important tools in the study of pathophysiology of glomerular diseases by providing valuable informations on the behavior of mesangial cells in normal and under various conditions.
It is necessary to obtain glomeruli for mesangial cell culture. Traditionally, isolation of glomeruli from renal cortesx has been done using repeated sieving method. But it is a very difficult and inconvinient procedure to perform. So authors
tried
iron
oxide particle perfusion method, a novel but more simple method for the isolation of glomeruli, instead of using repeated sieving method, the classical one, Authors also attempted to culture rabbit mesangial cells using acquired glomeruli and
confirm
the purification by observing gross morphology of cultured cells and patterns of immunofluorescent staining with various antibodies.
@ES The results were as follows:
@EN 1) Rabbit glomeruli were isolated with higher purity and rapidity by iron oxide particle perfusion method.
2) Cultured rabbit mesangial cells were slightly slender and polygonal in morphology in their early stage. As propagation of cell culture continued, they became longer and more slender with angled sharpen tips. Confluent mesangial cells gathered
and
made lumpy mass named as hill-lock appearance.
3) In immunofluorescent staining study, rabbit mesangial cells were negative to anti cytokeratin antibody, anti-factor VIII related antibody, anti-rabbit MHC antibody and antidesmin antibody but were positive to anti-=smooth muscle actin
antibody
and
anti-vimentin antibody.
4) Rabbit mesangial cells were successfully cultured and purified in 8 among 10 New Zealand White rabbits included in this study.
From the above findings, iron oxide particle perfusion method was thought to be an useful and c9nvinient method in the isolation of glomeruli from renal cortex for measangial cell culture.
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