The Growth Promoting Effect of Insulin-like Growth Factor-I(IGF-I) Purified from Human Serum F208 on the Rat Rib Chondrocytes / 대한소아내분비학회지
Journal of Korean Society of Pediatric Endocrinology
;
: 197-206, 1997.
Article
in Korean
| WPRIM
| ID: wpr-208021
ABSTRACT
PURPOSE:
The pathogenesis of short stature in growth hormone(GH) deficiency is believed to be based on the growth failure of growth plate chondrocytes by reduced growth hormone dependent insulin-like growth factor- I (IGF- I ) level in serum. Therefore, author studied the growth promoting effect of IGF- I purified from human serum on the chondrocytes, cultured from rat rib cartilage.METHODS:
Rat rib cartilage were treated with type II collagenase and hyaluronidase and were cultured in Ham's F-12 culture media containing 10% fetal calf serum. Growth promoting effect of IGF- I was measured by MTT dye by adding 20ng/ml IGF- I purified by protein-diol 120 column(YMC Co, Japan) from human serum, to 1*104 cultured chondrocytes separated into each of 96 well culture vessel.RESULTS:
1) When elution time of biotin labeled IGF- I by protein pak 300sw column was compared to elution time of standard molecular weight, IGF- I exists as large complex of 150Kd and small complex of 50Kd with free 7Kd form in serum before acid treatment. After acid treatment, IGF- I exists as small complex of 50Kd with free 7Kd form. 2) IGF- I purified from blood samples, as compared to genetic engineering product standard IGF- I , showed good parallelism in competition inhibition curve by purity analysis utilizing IGF- I antibody, and thus it is assumed that complex protein as inhibiting factor for purified IGF- I does not exist. Furthermore, complex protein was not present on the Western ligand method using biotin-labeled IGF- I ligand after purified IGF- I transferred to nitrocellulose paper following SDS-PAGE electrophoresis. 3)IGF- I of 20ng/ml showed 30% growth promoting effect, when rat rib chondrocyte culture with Dulbeco's modified Eagles medium(DMEM) is considered to show maximum growth promoting effect, while with pure culture medium, DMEM, showing minimum effect.CONCLUSIONS:
The results of this study suggest that IGF- I purified by this method assumes the role of growth promoting effect on the chondrocytes, and that the described method of radioimmuno assay of IGF- I also could effectively remove inhibiting protein complex, therefore allowing more accurate assay.
Full text:
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Index:
WPRIM (Western Pacific)
Main subject:
Ribs
/
Staphylococcal Protein A
/
Biotin
/
Growth Hormone
/
Cartilage
/
Genetic Engineering
/
Collagenases
/
Collodion
/
Chondrocytes
/
Eagles
Limits:
Animals
/
Humans
Language:
Korean
Journal:
Journal of Korean Society of Pediatric Endocrinology
Year:
1997
Type:
Article
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