Numerous bone cell culture models have been presented by the development of isolation and culturing techniques of cells. The culture of osteoblast-like cells of human origin with a specific osteoblastic phenotype has become an important experimental model in bone biology. Recently, it has become increasingly popular to utilize bone marrow cultures because these cultures are therefore thought to represent earlier stages of the osteoblast differentiation pathway. There is no report about culturing normal human osteoblast from oral and maxillofacial area. Primary cultured cells from oral and maxillofacial cancellous bone were analyzed by morphologic features, total DNA contents, ALP, osteocalcin and von Kossa staining positivity. The purpose of this study were to culture the cell population from oral and maxillofacial cancellous bone and to analyze the phenotypic expression of cultured normal human osteoblast by the bone marrow isolation technique. Growth curve of NHost showed about 45hrs of doubling time and about 70μ g/well of total DNA content. NHost showed spindle shaped cytoplasm with ovoid nucleus under preconfluency and after cellular differentiation, they formed irregular numerous nodules from stratified cellular layers under D medium. ALP activity was about 2 folds higher under control medium with 10nM 1,25(OH)2D3 than that under control medium. Osteocalcin expression was about seven folds higher under control medium with 100nM 1,25(OH)2D3 than that under control medium. Scattered mineralized nodules stained by von Kossa method were seen on the cellular layer under D medium. It suggested that NHost might be established from oral and maxillofacial area by characteristic cellular shape, ALP activity, osteocalcin expression and numerous mineralized nodules.

I. INTRODUCTION
 II. MATERIALS and METHODS
  1. Cell Culture Isolation Techniques
  2. Total DNA Assay
  3. Alkalin Phosphatase(ALP) Activity Assay
  4. Osteocalcin Assay
  5. von Kossa Mineralization Assay
  6. Statistics
 III. RESULTS
  1. Culture Characteristics
  2. ALP Activity
  3. Osteocalcin Secretion
  4. von Kossa Staining
 IV. DISCUSSIONS
 V. REFERENCES

• 문용식(Department of Oral Pathology, The Oral Aging Research Center, Dental College, Dankook University) | Yong Sik Moon
• 박경주(Department of Oral Pathology, The Oral Aging Research Center, Dental College, Dankook University) | Gyeong Ju Park
• 이종헌(Department of Oral Pathology, The Oral Aging Research Center, Dental College, Dankook University) | Chong Heon Lee correspondence