摘要
目的检测巴马香猪不同部位来源肋软骨细胞增殖、凋亡和迁移能力,探讨肋软骨细胞生物学行为的区域性差异。方法8个月龄巴马香猪6只,取其第6~8肋软骨,分为近肋骨端(A组)、中段(B组)、近胸骨端(C组),获得软骨细胞。CCK-8法检测各组细胞增殖能力,测量各组吸光度值,绘制曲线。克隆形成实验检测各组细胞自我更新的能力,以形成的集落数计算克隆形成率。血清饥饿诱导软骨细胞凋亡,TUNEL染色标记凋亡细胞,计数各组凋亡细胞数。在细胞划痕实验中通过相对面积法计算各组细胞迁移率。组间比较采用方差分析,以P<0.05为差异有统计学意义。结果CCK-8结果显示,3组细胞增殖差异具有统计学意义(F=5.719,P=0.014)。各组克隆形成率:A组为(18.33±0.47)%,B组为(12±0.82)%,C组为(14.33±0.94)%,3组比较差异有统计学意义(F=34.63,P<0.001)。各组凋亡细胞数:A组为(29.57±3.87)个/视野,B组为(97.8±21.99)个/视野,C组为(64.2±12.81)个/视野,3组比较差异有统计学意义(F=34.01,P<0.001)。各组细胞迁移率:24 h时A组为(14.07±3.99)%、B组为(18.41±3.31)%、C组为(15.08±4.64)%,3组比较差异无统计学意义(P>0.05)。结论体外培养时,巴马香猪近肋骨端来源的肋软骨细胞表现出更高的增殖、自我更新能力和对缺营养环境的耐受能力,而不同部位来源的肋软骨细胞单位时间内细胞迁移率无显著差异。
Objective To compare the proliferation,apoptosis and migration ability in vitro of porcine costal cartilage chondrocytes from different regions of the same rib.Methods The 6-8th ribs were harvested and divided into three groups,the rib proximal region group(group A),the middle region group(group B)and the sternum proximal region group(group C).Chondrocytes were isolated and cultured in vitro with the same culture media.Proliferation of chondrocytes was assayed by CCK-8 kit.The self-renewal ability was measured by colony-forming units assay.Apoptosis of chondrocytes was induced by serum starvation and detected by TUNEL kit.The cell migration rate was measured by the scratching test.Results CCK-8 results showed that there was significant difference in cell proliferation between the three groups(F=5.719,P=0.014).The colony formation rate of group A,B and C showed to be(18.33±0.47)%,(12±0.82)%and(14.33+0.94)%,respectively.The difference between the three groups was statistically significant.(F=34.63,P<0.001).The number of apoptotic cells of each group was 29.57±3.87 in group A,97.8±21.99 in group B,and 64.2±12.81 in group C.There was significant difference between the three groups(F=34.01,P<0.001).The migration rate of group A,B,and C showed to be(14.07±3.99)%,(18.41±3.31)%and(15.08±4.64)%within 24 hours,respectively,showing no significant difference(P>0.05).Conclusions when cultured in vitro,the costal chondrocytes from the proximal rib end showed higher proliferation,self-renewal capacity and tolerance to nutritionally deficient conditions,while the cell migration rate of the costal chondrocytes from different rib segments showed no significant difference.
作者
杨峥
何乐人
Yang Zheng;He Leren(First Center of External Ear Reconstruction,Plastic Surgery Hospital,Peking Union Medical College,Chinese Academy of Medical Science,Beijing 100144,China)
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2020年第4期440-446,共7页
Chinese Journal of Plastic Surgery
基金
中国医学科学院医学与健康科技创新工程项目(2017-I2M-1-007)首都特色临床应用研究(Z181100001718161)。
关键词
肋软骨
细胞增殖
克隆形成
细胞凋亡
细胞迁移
Costal cartilage
Proliferation
Colony-forming rate
Apoptosis
Migration
