退变腰椎间盘髓核细胞立体培养与单层培养的生物活性比较  

Comparison of biological potentiality of degenerated human lumbar disc cells between three-dimensional microcarrier culturing and monolayer culturing

作　　者：刘勇[1] 胡有谷[1] 宁斌[1] 

机构地区：[1]青岛大学医学院附属医院骨科山东省创伤骨科研究所,青岛市266003

出　　处：《中国脊柱脊髓杂志》2007年第5期376-379,I0003,共5页Chinese Journal of Spine and Spinal Cord

基　　金：国家自然科学基金资助(编号:30271318)

摘　　要：目的:比较体外单层培养和旋转微载体立体培养人退变腰椎间盘髓核细胞的生物学活性指标,探讨更加有效的椎间盘髓核细胞体外培养方法。方法:对获取的腰椎间盘突出症患者的24个椎间盘按年龄分为A组(20～25岁)、B组(26～30岁)、C组(36～45岁)及D组(>45岁),分别利用酶消化法进行单层细胞培养和旋转微载体立体培养系统进行立体培养,观察细胞生长形态,检测细胞生长曲线及速度、细胞生长活性、细胞分裂指数及胶原含量。结果:单层培养的髓核细胞贴壁后为多角形或梭形,伸出伪足;立体培养的细胞在微载体上呈梭形或不规则形,呈立体状生长。立体培养的细胞生长速度较快,1周内两种培养方法各时间点及各组间比较均具有统计学意义(P<0.05)。立体培养的髓核细胞活性提高,随年龄增加细胞活性下降;指数生长期细胞分裂指数与单层培养相比有统计学意义(P<0.05);Ⅰ、Ⅱ型胶原含量与单层培养相比有统计学意义(P<0.05),A组分别与B组、C组及D组比较均有统计学意义(P<0.05)。结论:旋转立体培养人退变椎间盘髓核细胞的活性保持良好,较单层培养能够大量、优质收集种子细胞,可用于椎间盘组织工程中种子细胞的研究。Objective：To compare the biological potentiality of intervertebral disc cells between the three-dimension microcarrier culturing and the monolaycr culturing,and to find a more efficient way of in vitro disc cell culturing.Method：24 samples of human degenerated intervertebral discs obtained during surgical procedures were isolated and digested.All these samples were assigned according to age as group A （20-25 years old）,group B（26-30 years old）,group C（36-45 years old） and group D（〉45 years old）.Afier proliferation in monolayer culture,cells were seeded in a new three-dimension microcarrier stiring system.The cytobiology index containing cell curve,cell proliferating and cell growth activity were detected.The content of different type of collagen and proteoglycan in disc cells were analyzed quantetively,Result：It showed that three-dimension culturing could improve the cell growth rate and cell growth activity,There were significant difference with respect to the number of cells between monalayer culturing and three-dimension culturing after a week （P〈 0.05）.The indexes of cell division during growing period between monolayer culturing and three-dimension culturing showed significant difference （P〈0.05）,eompared group A to group D showed significant difference （P〈0.05）,too.Comparison of collagen type Ⅰ and typeⅡ between monolayer culture and three-dimensional culture showed significant difference （P〈0.05）.There were significant differeee between group A and the other three groups（P〈0.05）.Tbere were significant difference with respect to the amount of type Ⅰ ,Ⅱ eolagon between monolayer culturing and three-demension cultuing （P〈0.05）.Conclusion：Human disc cells cultured in three-dimension microcarrier stiring system could keep cell growth and the phenotype well as well as express more extraeellular matrix.The apoptosis rate of the cells in three-dimension culturing is lower than that in monolaver culturing which suggest that we can collect conside

关 键 词：椎间盘 退变 细胞培养 微载体 

分 类 号：Q813.1[生物学—生物工程] R681.5[医药卫生—骨科学]