Somatomedin-like peptide(s) isolated from fetal bovine cartilage (cartilage-derived factor): isolation and some properties.

Abstract

Communicated by H. A. Barker, August 10, 1981 ABSTRACT Fetal bovine cartilage contains a polypeptide(s) that has somatomedin-like effects on rat and rabbit costal chondrocytes in culture. This factor, named the cartilage-derived factor, was extracted from fetal bovine cartilage, fractionated with acetone, and purified by gel fitration on Toyopearl HW 55-F in 4 M guanidine hydrochloride, preparative isoelectric focusing, and subsequent gel filtration on Toyopearl HW 55-F in 1 M formic acid. The resulting preparation, which focused in the neutral pH region and eluted from a Toyopearl column in a fraction with apparent Mr 10,000-11,000, appeared homogenous by NaDodSO4 gel electrophoresis. The purified preparation markedly enhanced not only proteoglycan synthesis but also DNA synthesis in rabbit costal chondrocytes and, on a protein basis, it was 1000 times more active than insulin and 1,000,000 times more active than fetal calf serum in stimulating proteoglycan synthesis.

Local bone growth factors

“…The effect of BDGF is not specific for bone since it stimulates the growth of cartilage and fibroblasts. The effect on cartilage is similar to that of a cartilage-derived factor [6] and that on fibroblasts is shared by TGFs, but highly purified BDGF does not appear to contain transforming activity.…”

Effect of cartilage-derived factor on DNA and protein synthesis in cultured rat calvariae

“…Our results also suggest that CDF did not act by inhibiting collagen degradation, since only a small and similar proportion of the newly labeled macromolecular collagen was released in both control and treated cultures. The doses of CDF that increased bone DNA, collagen, and noncollagen protein synthesis were higher than those required to stimulate DNA and proteoglycan synthesis in chondrocyte cultures [5], indicating that cartilage is more sensitive to CDF than bone. This is not surprising in view of the cartilage origin of CDE Further, CDF was effective on bone at doses similar to those required by a bone-derived growth factor and by Multiplication Stimulating Activity to increase collagen and DNA synthesis in calvarial cultures [1,17].…”

“…In addition, a growth factor with an approximate molecular weight of 10,000 has been isolated from bovine fetal cartilage. This factor, named cartilagederived factor (CDF), stimulates DNA and proteoglycan synthesis in cultured chondrocytes [5] but its effects on bone are as yet unknown.The present studies were undertaken to study further the relationship between bone and cartilage growth by examining the effects of partiaUypurified CDF on bone formation. For this purpose, the effects of CDF on bone DNA, collagen, and noncollagen protein synthesis were studied in cultures of rat calvariae.…”

Purification and characterisation of a growth factor from porcine bone

“…Endogeneous bone growth factors of possible relevance are produced from organ culture or extracted from bone matrix and include: bone-derived growth factor, a 11.6-kDa protein produced by cultured fetal rat calvaria [l, 51; transforming growth factor-p, a 25-kDa protein recently demonstrated by the same organ culture system [5, 61; cartilagederived factor, a substance like somatomedin C and insulin growth factor I [7]; cationic cartilage-derived growth factor, a 19-kDa protein like fibroblast growth factor [S]; human skeletal growth factor, a 9 -12-kDa form now recognized as being like insulin growth factor I1 [9, lo]; bone morphogenetic protein, an 18-kDa protein isolated from bovine bone matrix [ll]; osteogenin, an 18.5-kDa bovine protein with bone-inducing activity [3]; a 22-kDa mitogenic factor from rat bone [12]; and a 26-kDa cartilage induction factor A from bovine bone 1131, now recognized to be identical to transforming growth factor [14], and cartilage induction factor B and [l, 5, 6, 121, ox [3, 11, 19, 211 and human [9, 101. The present report describes a growth factor, isolated from porcine bone matrix, using the mitogenic response of osteoblast-progenitor cell populations of embryonic chick calvaria.…”

“…The osteoprogenitor cell proliferation process is well known and seen in the reaction of periosteum and endosteum to injury, diet, vitamins and hormones. The process of induced cell differentiation of niesenchymal cells has been observed from quantitative measurement of bone formed in response to implants of either bone matrix or purified proteins in extraskeletal and intraskeletal sites [2-41. Endogeneous bone growth factors of possible relevance are produced from organ culture or extracted from bone matrix and include: bone-derived growth factor, a 11.6-kDa protein produced by cultured fetal rat calvaria [l, 51; transforming growth factor-p, a 25-kDa protein recently demonstrated by the same organ culture system [5, 61; cartilagederived factor, a substance like somatomedin C and insulin growth factor I [7]; cationic cartilage-derived growth factor, a 19-kDa protein like fibroblast growth factor [S]; human skeletal growth factor, a 9 -12-kDa form now recognized as being like insulin growth factor I1 [9, lo]; bone morphogenetic protein, an 18-kDa protein isolated from bovine bone matrix [ll]; osteogenin, an 18.5-kDa bovine protein with bone-inducing activity [3]; a 22-kDa mitogenic factor from rat bone [12]; and a 26-kDa cartilage induction factor A from bovine bone 1131, now recognized to be identical to transforming growth factor [14], and cartilage induction factor B and The present report describes a growth factor, isolated from porcine bone matrix, using the mitogenic response of osteoblast-progenitor cell populations of embryonic chick calvaria. The growth factor was purified by heat and acid treatment, and chromatography on hydroxyapatite, gel-filtration under dissociative conditions and reverse-phase HPLC.…”