1.Odontogenesis-related gene expression during in vitro culture of tooth germ cells
Jiaojiao BA ; Huiyu HE ; Yang HU ; Mei MA ; Xiangzhen HAN
Chinese Journal of Tissue Engineering Research 2014;(2):193-198
BACKGROUND:Some studies have indicated that different genes in tooth germ tissue play a role at different time, contributing to tooth germ development.
OBJECTIVE:To observe the expressions of dentin matrix protein 1, enamel protein, col agen I and homeobox gene 1 at different stage of in vitro culture of tooth germ cells.
METHODS:RNA from tooth germ cells was extracted at days 1, 3, 6 of in vitro culture. After reverse transcription, real-time quantitative PCR detection was adopted to measure relative expression of dentin matrix protein 1, enamel protein, col agen I and homeobox gene 1 mRNA.
RESULTS AND CONCLUSION:Dentin matrix protein 1, enamel protein, and col agen I mRNA expressions increased with culture time, and reached the peak at day 3 (P<0.05), whereas homeobox gene 1 mRNA decreased with culture time (P<0.05).
2.Recombinant lentiviral vector transfected sheep bone marrow mesenchymal stem cells and osteogenic gene expression changes
Xiangzhen HAN ; Huiyu HE ; Yang HU ; Jiaojiao BA ; Huanhuan WANG ; Xue MI
Chinese Journal of Tissue Engineering Research 2014;(6):821-828
METHODS:Lentiviral vectors carrying bFGF and BMP-2 were constructed to transfect sheep bone marrow mesenchymal stem cells. cells were divided into four groups:bFGF group, BMP-2 group, co-transfection group BACKGROUND:Basic fibroblast growth factor (bFGF) can promote the proliferation of bone marrow stromal cells, and bone morphogenetic protein-2 (BMP-2) has an important significance in the induction of new bone formation.
OBJECTIVE:To analyze the effects of bFGF, BMP-2 and their co-transfection on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and to compare the relative expressions of col agen I, osteocalcin and osteopontin before and after celltransfection, thereby providing theoretical implications for seed cells in the construction of tissue-engineered bone.
and control group. The RNA was extracted using real-time quantitative PCR to detect mRNA levels of col agen I, osteocalcin, and osteopontin.
RESULTS AND CONCLUSION:Significant difference in non-specific osteogenic gene expressions was found among the four groups (P<0.05). bFGF and BMP-2 showed an interaction (P<0.05). Expressions of col agen I and osteocalcin in the co-tranfection group were higher than those in the other three groups (P<0.05), but osteopontin expression exhibited no difference (P>0.05). In vitro experiments showed that the relative expression of col agen I, osteocalcin and osteopontin were higher in the co-transfection group, indicating the cells from the co-transfection group have strongest osteogenic capacity that are suitable for seed cells for bone tissue engineering.