二维和三维培养的人皮肤成纤维细胞中生长因子mRNA表达的比较研究

COMPARISON OF GROWTH FACTOR mRNA EXPRESSIONS IN HUMAN SKIN FIBROBLASTS BETWEEN TWO AND THREE-DIMENSIONAL CULTURE SYSTEMS

探讨经二维或三维培养的人皮肤成纤维细胞在细胞因子表达能力方面是否有差异。以平面培养皿作为成纤维细胞二维培养系统 ,用胶原凝胶作为成纤维细胞三维培养系统 ,分别提取二维和三维培养的人皮肤成纤维细胞总RNA ,采用RT PCR方法检测两种培养系统中血管内皮细胞生长因子(VEGF)和碱性成纤维细胞生长因子 (bFGF)mRNA的表达。结果显示 ,虽然二维和三维培养的人皮肤成纤维细胞形态不同 ,但二维和三维培养的成纤维细胞均有VEGF/bFGF设计大小的条带显示 ,且两者条带相对灰度无明显差异。

To compare the growth factors mRNA expressions in human skin fibroblasts cultured in two (2D) or three dimensional systems (3D). Human skin fibroblasts were isolated and cultured in collagen jelly (3D) or the regular culture plates (2D). After 72 hours, the total RNA of fibroblasts were extracted. Specific oligonucleotide primers for the growth factor VEGF and bFGF were synthesized and reverse transcriptase polymerase chain reaction (RT PCR) was utilized to amplify the mRNA of those growth factors. RT PCR productions were separated by electrophoresis in 1% wt/vol agarose gels and photographed. Semi quantitative data were statistically analyzed. Although the morphology of fibroblasts in 3D culture was different from that in 2D culture, the RT PCR analysis revealed the same expression and concentration of VEGF mRNA and bFGF mRNA in human skin fibroblasts cultured in different systems.