دستيابي به كشت سه بعدي سلول هاي استيو بلاست مشتق ازكالوارياي انساني در ايران

Access to a Three Dimentional Osteoblasts Culture in Iran

The problem of bone defects during treatments of bone lesions, especially cranial trauma, and also during craniotomy operations indicates isolation of osteoblasts and culturing them for filling these bone effects. Bone tissue specimens were obtained from 6 patientsʹ carvaria undergoing craniotomy surgery operations in Al-zahra teaching hospital and transferred to lab. These specimens were cut into small pieces measuring lxl mm and put in Petri dishes having culture medium and then transferred to the incubator. The mean interval of observed cellular outgrowth was 9-10 days. After 2 weeks, the cells in cultures reached confluencey. First passage cells were detached from Petri dishes, using Trypsin EDTA, and were divided in two portions. One portion was used for monolayer osteoblast culture, and the other, was added to alginate gel. The activity of these cells was tested by Alkalin phosphatase, Von kossa and Azan staining. Cell viability was also tested by Trypan blue staining. The morphology of osteoblasts in monolayer cultures appeared fusiform and fibroblastic, while in the alginate beads, cells appearance was round. Alkaline phosphatase staining method confirmed the presence of osteoblasts. Von kossa and Azan staining demonstrated, mineralized and organised matrix in both groups. The number of harvested cells in day 1 and day 21 were significantly different in each group (P < 0.001). In addition, there was significant difference in the mean of cells number and viability between monolayer cultures and alginate beads in day 21 (P< 0.001). This study showed that osteoblasts can come out from bones in dish culture without using any enzyme; and sodium Alginate gel supports cell viability and proliferation of osteoblasts cells significantly in comparison with the monolayer culture.