[3D Collagen Hydrogel Culture of Rat Calvarial Osteoblasts].
OBJECTIVE
To establish a collagen hydrogel three-dimensional culture model with rat calvarial osteoblasts (ROBs).


METHODS
ROBs were obtained through enzyme digestion of segregated neonatal SD rat skull. The collagen hydrogel three-dimensional culture model was established by mixing ROBs with different concentrations of type I rat tail collagen (collagen concentration of 1, 2, 3 mg/mL), DMEM medium and NaOH under adjusted PH and a temperature of 37 degrees C. Cell viability and activity were detected by FDA/PI staining and CCK-8 3 d after cell culture. The optimal culture method of 3D collagen hydrogel was identified. Cell distribution was observed using scanning electron microscopy and HE staining.


RESULTS
ROBs collagen was formed firmly at 2 mg/mL, which had significantly higher levels of cell viability and activity than those at 1 mg/mL and 3 mg/mL. Scanning electron microscopy and HE staining showed that cells under the 2 mg/mL collagen culture system adhered with collagen tightly and distributed homogeneously.


CONCLUSIONS
A collagen hydrogel 3D culture model was established successfully by mixing ROBs with collagen at 2 mg/mL.
Yan-fang
YF
Xie
Wen-gui
WG
Shi
Jian
J
Zhou
Yu-hai
YH
Gao
Ming-gang
MG
Wang
Ke-ming
KM
Chen
D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival; D024042:Collagen Type I / Q000737:chemistry*; D020100:Hydrogels / Q000737:chemistry*; D010006:Osteoblasts / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012886:Skull / Q000166:cytology
D024042:Collagen Type I; D020100:Hydrogels