United States Patent
US Patent 6358737: Osteocyte cell lines
US 6358737 · granted 2002-03-19
Abstract
The present invention relates to a method of producing osteocyte cell line in various stages of differentiation. Such cell line remains stable after more than 20 passages. The osteocyte has a stellate shape with dendritic processes and expresses high level of osteocalcin. More specifically, it provides a method of production for cultured osteocytes of various differentiation stages. Furthermore, it relates to osteocyte cell line, and more specifically cultured osteocyte. The invention also relates to a method for the production of monoclonal antibodies using such cultured osteocytes and further relates to hybridomas and monoclonal antibodies which recognize an osteocyte-specific antigen. Finally, the invention relates to a method of screening for modification factors and binding factors for osteocytes.
| Patent Number | 6358737 |
|---|---|
| Title | Osteocyte cell lines |
| Filed | 1998-09-11 |
| Granted | 2002-03-19 |
| Inventor(s) | Bonewald; Lynda F, Kato; Yoichi |
| Number of Claims | 104 |
Abstract
The present invention relates to a method of producing osteocyte cell line in various stages of differentiation. Such cell line remains stable after more than 20 passages. The osteocyte has a stellate shape with dendritic processes and expresses high level of osteocalcin. More specifically, it provides a method of production for cultured osteocytes of various differentiation stages. Furthermore, it relates to osteocyte cell line, and more specifically cultured osteocyte. The invention also relates to a method for the production of monoclonal antibodies using such cultured osteocytes and further relates to hybridomas and monoclonal antibodies which recognize an osteocyte-specific antigen. Finally, the invention relates to a method of screening for modification factors and binding factors for osteocytes.
Claim 1
A method for the production of an osteocyte cell line, said cell line having the following properties: available to passage more than 20 times, a stellate shape withdendritic processes and expresses increased levels of osteocalcin compared to mouse osteoblast or osteoblast-like cell lines, and said method comprising the steps of: (a) isolating bones from a transgenic mouse whose genome contains an osteocalcin promoter driven T-antigen transgene and preparing bone pieces from the bones; (b) digesting the bone pieces with collagenase solution and harvesting cells into fetal and adult calf serum supplemented medium; and (c) plating the harvested cells and isolating said cell line by selecting a single colony.
Claims
104 totalA method for the production of an osteocyte cell line, said cell line having the following properties: available to passage more than 20 times, a stellate shape withdendritic processes and expresses increased levels of osteocalcin compared to mouse osteoblast or osteoblast-like cell lines, and said method comprising the steps of: (a) isolating bones from a transgenic mouse whose genome contains an osteocalcin promoter driven T-antigen transgene and preparing bone pieces from the bones; (b) digesting the bone pieces with collagenase solution and harvesting cells into fetal and adult calf serum supplemented medium; and (c) plating the harvested cells and isolating said cell line by selecting a single colony.