ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of co-expression of bone morphogenetic protein 2 (BMP2) and Sox9 on chondrogenic differentiation of mesenchymal stem cells (MSCs) in vitro and provide experimental evidence for tissue engineering of cartilage.</p><p><b>METHODS</b>Mouse embryonic bone marrow MSC C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP2, Sox9 and green fluorescent protein (GFP) for 3-14 days, with cells infected with the adenovirus carrying GFP gene as the control. The mRNA expression of the markers of chondrogenic differentiation, including collagen type II (Col2a1), aggrecan (ACAN), and collagen type X (Col10a1), were determined by real-time PCR. Alcian blue staining was used for quantitative analysis of sulfated glycosaminoglycan in the cellular matrix. The expression of Col2a1 protein was assayed by immunohistochemical staining and Western blot analysis.</p><p><b>RESULTS</b>Adenovirus-mediated BMP2 expression induced chondrogenic differentiation of C3H10T1/2 cells. Overexpression of Sox9 effectively enhanced BMP2-induced expression of the chondrogenic markers Col2a1, aggrecan and Col10a1 mRNAs, and promoted the synthesis of sulfated glycosaminoglycan and Col2a1 protein in C3H10T1/2 cells.</p><p><b>CONCLUSION</b>Co-expression of BMP2 and Sox9 can promote chondrogenic differentiation of MSCs in vitro, which provides a new strategy for tissue engineering of cartilage.</p>
Subject(s)
Animals , Humans , Mice , Bone Morphogenetic Protein 2 , Genetics , Metabolism , Cartilage , Cell Biology , Cell Differentiation , Cells, Cultured , Chondrocytes , Cell Biology , Mesenchymal Stem Cells , Cell Biology , Metabolism , SOX9 Transcription Factor , Genetics , Metabolism , Tissue EngineeringABSTRACT
Objective To study the role of Cysteine-rich 61 ( Cyr61/CNN1) in repair of combined injury by observing whether exogenous CCN1 promotes the proliferation and migration of radiation-injured L929 cells. Methods A radiation model of L929 cells was induced by ? ray at a dose of 4 Gy. The irradiated L929 cells were cultured in a medium containing 2 ml adenovirus plasmids of CCN1 or RFP at 37 ℃ in an atmosphere containing 5% CO2,which served as a CCN1 group and a RFP group,respectively. Irradiated L929 cells cultured in a blank control medium served as a blank control group. Effects of CCN1 on proliferation and migration of radiation-injured L929 cells were detected by MTT assay,plate colony formation assay,cell cycle analysis,and scratch test of wound healing. Results The proliferation and colony formation rates of L929 cells cultured in a medium containing CCN1 were significantly higher than in RFP and control groups [colony formation rate:( 34. 4 ?3. 6) % vs ( 24. 5 ?2. 9) % and ( 29. 5 ?3. 5) %,P
ABSTRACT
Objective To observe the effects of radiation on the growth and expression of cysteine-rich 61(Cyr61/CCN1) of L929 cells and investigate the relationship between CCN1 expression and radiation injury.Methods L929 cells were cultured and divided into 2 groups,cells irradiated with 4 Gy ?-irradiation as radio-group and untreated cells as control group.The cell proliferation was measured by MTT assay and plate colony formation testing.Flow cytometry was utilized to quantify the cell cycle distribution.CCN1 expression at protein and mRNA levels were determined by immunocytochemistry(ICC) and RT-PCR respectively.Results Significant inhibition of proliferation(P