CAJ | 학술논문

背景:目前尚未见成熟的成年人退行性变椎间盘的细胞培养模型报道,而此模型为建立椎间盘组织工程的细胞学基础.目的:建立成人椎间盘细胞培养模型,奠定椎间盘组织工程研究的细胞学基础.设计及地点:对比观察的细胞学实验,在山东省创伤骨科研究所完成.材料:标本来源于青岛大学附属医院骨科5例确诊为退行性椎间盘病变患者.L2..椎间盘1例,L4～5椎间盘3例,L5～S1椎间盘1例.方法:对5例取自腰椎间盘突出症退变椎间盘的手术标本,采用系列酶消化法细胞培养和组织块细胞培养两种方法,应用HAMF12培养基加体积分数为10%和20%的胎牛血清分别进行了细胞培养,均获得了传代,并对培养细胞进行了光镜和电镜的形态学观察.主要观察指标:①细胞生长状况.②光镜下细胞形态观察.③透射电镜超微结构观察.结果:酶消化法和组织块法均能获得大量的退变椎间盘细胞,并成功地进行细胞传代.培养细胞的分泌物质可能对细胞的生长起一定的限制作用,去除该物质后细胞可迅速增生.但在细胞生长及增殖过程中,体积分数为20%胎牛血清下细胞增殖速度明显高于体积分数为10%胎牛血清时的速度.全部退变椎间盘标本中均发现了脊索细胞.结论:成人退变椎间盘可以用来获得椎间盘组织工程所需的细胞.
배경:목전상미견성숙적성년인퇴행성변추간반적세포배양모형보도,이차모형위건립추간반조직공정적세포학기출.목적:건립성인추간반세포배양모형,전정추간반조직공정연구적세포학기출.설계급지점:대비관찰적세포학실험,재산동성창상골과연구소완성.재료:표본래원우청도대학부속의원골과5례학진위퇴행성추간반병변환자.L2..추간반1례,L4～5추간반3례,L5～S1추간반1례.방법:대5례취자요추간반돌출증퇴변추간반적수술표본,채용계렬매소화법세포배양화조직괴세포배양량충방법,응용HAMF12배양기가체적분수위10%화20%적태우혈청분별진행료세포배양,균획득료전대,병대배양세포진행료광경화전경적형태학관찰.주요관찰지표:①세포생장상황.②광경하세포형태관찰.③투사전경초미결구관찰.결과:매소화법화조직괴법균능획득대량적퇴변추간반세포,병성공지진행세포전대.배양세포적분비물질가능대세포적생장기일정적한제작용,거제해물질후세포가신속증생.단재세포생장급증식과정중,체적분수위20%태우혈청하세포증식속도명현고우체적분수위10%태우혈청시적속도.전부퇴변추간반표본중균발현료척색세포.결론:성인퇴변추간반가이용래획득추간반조직공정소수적세포.
BACKGROUND: There are no reports about adult degenerative disc cell culture model currently. This establishment of disc cell culture is the cellular basis of intervertebral disc tissue engineering. OBJECTIVE: To establish cell culture models from human degenerative intervertebral disc, and to settle the cytology base of disc tissue engineering. DESIGN AND SETTING: The controlled observational experiment with regard to cytology was performed in Shandong Institute of Orthopaedics and Traumatology. MATERIALS: Five degenerative intervertebral disc patients were recruited from Department of Orthopaedics, Affiliated Hospital of Qingdao University Medical College, including one case in L2-3, three cases in L4-5 and one case in L5-S1. METHODS: Five samples were taken from the operational disectomy for lumbar disc herniation, and these samples were cultured using enzymatic digestion cell culture and tissue cell culture method differently. All the samples were cultured with HAMF12 medium with the addition of 10% and 20% fetal bovine serum. The morphology of the cultured cell was observed with Giernsa staining and transmission electron microscopy respectively. MAIN OUTCOME MEASURES: ①Cellular growth; ②Morphology of the cultured cells;③Ultrastructure Of the cultured cells. RESULTS: By means of enzymatic digestion cell culture and tissue cell culture method, numerous degenerative lumbar disc cells were obtained and successfully subcultured. The cell secretions around the cultured cells may influence the growth of these cells. Destroying the secretions, the cultured cells could be highly proliferated. The degenerative intervertebral disc cells were proliferated vigorously in HAMF12 medium with 20% fetal bovine serum compared with that with 10% fetal bovine serum. The notochordal cell was observed in all the specimens. CONCLUSION: The adult degenerative disc can be used to obtain the required cell for tissue engineering.