成人退变椎间盘细胞微载体培养的形态学观察

MICROCARRIER CULTURE AND MORPHOLOGICAL OBSERVATION OF HUMAN DEGENERATIVE INTERVERTEBRAL DISCS

①目的建立成人退变椎间盘细胞的体外三维培养模型,并观察其形态学变化。②方法对8例取自椎间盘疾病病人退变椎间盘的手术标本,采用系列酶消化法对其进行了体外细胞培养,并将细胞转移至微载体旋转培养系统中进行培养,利用倒置相差显微镜进行形态学研究,并对细胞涂片进行染色观察。③结果退变椎间盘细胞在三维微载体旋转培养系统中可以获得良好的生长并大量的增生,培养的细胞具有退变的特性。④结论成人退变椎间盘细胞体外立体培养成功,退变细胞能够快速大量增殖,为成功体外构建组织工程学椎间盘奠定了基础。

Objective To establish an in-vitro three-dimension culture model of degenerative intervertebral disc cells and to observe the morphology of them. Methods Human degenerated intervertebral disc cells were isolated from eight samples of disc tissue, including four of protruded discs, two of ruptured discs, one of isolated disc and one of disc with scoliosis during surgical procedures. All the samples were cultured with HAMF 12 medium, and after proliferation in monolayer culture, the cells were cultured in a new three-dimensional microcarriers system. The morphology of these ceils was observed with TEM. Results In the new three-dimensional environment, the cells grew well and proliferated vigorously. The cultured cells were found to be of degenerative characteristics. Conclusion An in-vitro stereo-culture of adult degenerative intervertebral disc ceils is succeeded. The cells can massively and rapidly multiply, which settles a base for constructing intervertebral disc engineering.