Production of functional human bone morphogenetic protein-2 using a baculovirus/Sf-9 insect cell system.
A clone of a human Bone Morphogenetic Protein-2 (hBMP-2) cDNA was obtained from a cDNA library established from human dental pulp cells. After subcloning hBMP-2 cDNA into Autographa californica nuclear polyhedrosis virus, the recombinant baculovirus was transfected to Sf-9 cells. Immuno-reactive recombinant hBMP-2 (rhBMP-2) was detected by a polyclonal antibody against Xenopus BMP-2 in the transfected insect cells but not in the culture media. Three days after treatment with the lysate of the transfected Sf-9 cells, increase in alkaline phosphatase activity of a murine stromal cell line, ST2, was detected. Subcutaneous implantation of rhBMP-2 produced in the insect cells induced formation of cartilage, bone and bone marrow in the rats. The present data indicated that the rhBMP-2 preparation produced in the insect Sf-9 cells had a comparable activity to that produced in mammalian cells.
Maruoka
Y
Y
First Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Tokyo Medical and Dental University, Japan.
Oida
S
S
Iimura
T
T
Takeda
K
K
Asahina
I
I
Enomoto
S
S
Sasaki
S
S
D000469:Alkaline Phosphatase / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line / Q000821:virology; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D003782:Dental Pulp / Q000166:cytology; D015967:Gene Expression Regulation, Viral; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D000077183:Occlusion Body Matrix Proteins; D010012:Osteogenesis / Q000187:drug effects; D011401:Promoter Regions, Genetic; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D018411:Spodoptera / Q000166:cytology* / Q000821:virology; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics; D015678:Viral Structural Proteins
D018076:DNA, Complementary; D000077183:Occlusion Body Matrix Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D000469:Alkaline Phosphatase