| 摘要: |
| 目的:比较成人正常和突出椎间盘髓核细胞体外培养的形态及生长动力学差异.方法:从成人正常和突出髓核组织分离培养髓核细胞,分别取传1、3、5代细胞作为研究对象,光镜和电镜下观察细胞形态,并对细胞生长速率、MTT摄取、最大生长密度、分裂指数、克隆形成率等生长动力学指标进行比较研究.结果:体外培养的突出髓核细胞与正常髓核细胞相比,形态学上可见突出髓核细胞排列较紊乱,极性消失,胞质内粗面内质网轻度扩张.在接种相同的细胞密度和相同的培养条件下,成人正常和突出髓核细胞的生长速率差异在传1代时为T＝52.5(P>0.05),传3、5代分别为T＝72.0(P<0.01)、T＝67.5(P<0.05);二者的最大生长密度、MTT摄取、分裂指数、克隆形成率在传1代时分别为7.28±0.23和7.41±0.17、0.186±0.015和0.173±0.008、(25.36±2.14)‰和(23.13±2.55)‰、(52.4±2.8)％和(47.7±2.6)％,差异均无显著性(P>0.05),传3代时分别为6.96±0.19和5.48±0.22、0.154±0.018和0.110±0.009、(22.52±2.16)‰和(16.68±2.35)‰、(48.7±3.3)％和(40.7±2.5)％,传5代时分别为2.42±0.13和0.98±0.14、0.085±0.006和0.050±0.007、(15.61±1.96)‰和(9.34±2.12)‰、(40.2±2.3)％和(28.34±2.9)％,传3、5代时差异均有统计学意义(P<0.05).结论:成人正常和突出椎间盘的髓核细胞在体外培养状态下,突出组细胞在细胞形态学上与正常对照组存在一定差异,在生长动力学方面,突出组细胞的持续增殖能力明显低于正常髓核细胞组,更容易出现老化现象. |
| 关键词: 椎间盘、髓核、细胞,培养的 |
| DOI：10.3724/SP.J.1008.2006.00479 |
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| 基金项目:国家自然科学基金（30370389）. |
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| Comparison of morphology and growth kinetics of adult normal and herniated nucleus pulposus cells cultured in vitro |
| 张荣峰,张超,阮狄克,ZHANG Rong-feng,ZHANG Chao,RUAN Di-ke |
| () |
| Abstract: |
| Objective：To investigate the morphology and growth kinetics of the cells derived from the adult normal and herniated nucleus pulposus tissues. Methods： The cells derived from the adult normal and herniated nucleus pulposus were cultured in vitro. Passage 1,3,5 cells were chosen for light microscope and electron microscope observation. The cell growth rate, MTT value, maximal growth density, cellular mitotic index, cloning efficiency, and other parameters were observed in normal and herniated nucleus pulposus and the results were compared. Results： Compared with adult normal nucleus pulposus cells, the cells isolated from herniated nucleus pulposus were irregularly arranged and lost polarization, with rough endoplasmic reticulum slightly dilated. Under the same cellular density and culture conditions, the growth rate of passage 1 cells from adult normal and herinated nucleus pulposus had no significant difference, but the differences of passage 3 cells（P〈0.01） and passage 5 cells （P〈0.05） were statistically significant. The maximal growth density, MTT value, cellular mitotic index, and cloning efficiency of passage 1 cells of adult normal and herinated nucleus pulposus were respectively 7.28 ±0.23 and 7.41 ± 0. 17,0. 186 ± 0. 015 and 0. 173±0. 008, （25.36±2. 14）‰ and （23. 13±2.55）‰,and （52.4±2.8）% and （47.7±2.6）% （all P〉0.05）; those of passage 3 cells were respectively 6.96 ± 0.19 and 5.48± 0.22, 0.154 ± 0.018 and 0.110 ± 0. 009, （ 22.52 ± 2.16） ‰ and （16.68±2.35）‰, and （48. 7±3.3）% and （40.7±2.5）±;and of passage 5 cells were respectively 2.42±0. 13 and 0.98± 0.14, 0.085±0. 006 and 0. 050±0. 007, （15.61± 1.96）%o and （9.34±2. 12）%0, and （40.2±2.3） % and （28.34± 2.9） %. There were significant differences between the parameters of passage 3 and 5 cells（P〈0.05）. Conclusion： The adult normal and herinated nucleus pulposus cells are different in cellular morphology when cultured in vitro. The continuous proliferation ability of the herniated nucleus pulposus cells is obviously decreased than that of the adult normal control group,and aging is more easi- ly seen in the herniated ones |
| Key words: intervertebral nucleus pulposus cells,cultured |
