论文部分内容阅读
目的无血清培养条件下,研究不同年龄人表皮细胞(humanepidermalcells,hECs)的体外扩增规律,比较不同年龄hECs原代细胞表型。方法临床收集50例包皮环切术中丢弃的皮肤组织,包括中年组(A,35～52岁,11例)、青年组(Y,18～27岁,17例)、儿童组(C,2～12岁,22例)。以Dispase和胰酶(tripsin)-EDTA分离,收集hECs并进行体外培养。每个年龄组随机选取5例,各代细胞计数,计算最大扩增倍数,并统计分析。收集原代细胞测量平均直径,记录克隆形成率,流式细胞仪(FACS)检测细胞K19阳性百分率。结果中年、青年、儿童组包皮来源的表皮细胞最大扩增倍数分别为(719±65)、(730±47)、(729±56)倍,各组间无统计学差异(P>0.05)。全体随机标本平均可扩增(728±53)倍,达到最大扩增量所需平均时间为(31±6)d。中年、青年、儿童组,表皮细胞平均直径分别为(3.44±0.42)μm,(3.54±0.46)μm,(3.55±0.37)μm,各组间无统计学差异(P>0.05);克隆形成率分别为(36.5±5.3)%,(36.9±6.8)%,(36.4±5.1)%,各组间无统计学差异(P>0.05);FACS检测中K19阳性细胞百分率分别为,(42.55±4.61)%,(42.47±6.58)%,(41.77±4.73)%,各组间无统计学差异(P>0.05)。结论无血清培养hECs的体外扩增规律与年龄因素无关,不同年龄hECs最大扩增能力取决于原代细胞表型?
Objective To investigate the in vitro expansion of human epidermal cells (hECs) of different ages under serum-free culture conditions and to compare the phenotypes of hECs primary cells at different ages. Methods Fifty cases of skin tissue discarded during circumcision were collected, including middle age group (A, 35-52 years old, 11 cases), young group (Y, 18-27 years old, 17 cases), children group (C, 2 to 12 years old, 22 cases). The cells were separated by Dispase and tripsin-EDTA, hECs were harvested and cultured in vitro. In each age group, 5 cases were randomly selected, each generation of cells count, calculate the maximum fold, and statistical analysis. The primary cells were collected to measure the average diameter, and the rate of colony formation was recorded. The positive percentage of K19 cells was detected by flow cytometry (FACS). Results The maximum fold expansion of preputial cells derived from the foreskin of middle age, youth and children was (719 ± 65), (730 ± 47) and (729 ± 56) times, respectively. There was no significant difference between the groups (P> 0.05) . The average of all random samples can be amplified (728 ± 53) times, the average time required to reach the maximum amplification was (31 ± 6) d. The average diameters of middle-aged, young, children and epidermis were (3.44 ± 0.42) μm and (3.54 ± 0.46) μm and (3.55 ± 0.37) μm, respectively. There was no significant difference between the groups (P> 0.05) (36.5 ± 5.3)%, (36.9 ± 6.8)% and (36.4 ± 5.1)%, respectively. There was no significant difference among the groups (P> 0.05). The percentage of K19 positive cells in FACS was (42.55 ± 4.61%, 42.47 ± 6.58% and 41.77 ± 4.73%, respectively. There was no significant difference among the groups (P> 0.05). Conclusion The in vitro expansion of serum-free cultured hECs has nothing to do with age, and the maximum ability of hECs at different ages depends on the phenotype of primary cells.