摘要
目的:探讨正常皮肤和瘢痕疙瘩组织成纤维细胞在体外培养情况下其生物学特性的差异,以及与细胞冻存和复苏的方法及其应用价值。方法:对8例正常皮肤和8例瘢痕疙瘩组织标本分别进行体外成纤维细胞的原代培养、传代培养,观察培养的成纤维细胞形态,并作生长曲线比较细胞增殖情况。选取处于对数生长期的培养细胞梯度降温后置入液氮(-196℃)中保存4～6个月,进行细胞复苏培养,以2%台盼蓝确定细胞活力,并观察复苏的成纤维细胞形态学状况。结果:体外培养的正常皮肤和瘢痕疙瘩组织成纤维细胞形态和生长曲线基本相同,但瘢痕疙瘩原代成纤维细胞萌出生长较正常皮肤快,生长融合后成纤维细胞排列紊乱,极性消失,有明显的交叉重叠现象;体外培养的成纤维细胞,经冷冻保存4～6个月后,复苏率超过80%,细胞形态无明显改变。结论:体外培养的正常皮肤和瘢痕疙瘩组织成纤维细胞形态与生长特性基本相同。瘢痕疙瘩成纤维细胞可成功冻存和复苏。
Objective To investigate the difference of biological characteristics and growth status between culturing fibrablasts of normal skin and keloid in vitro,the viability of revivificated cells and the change of morpHology after revivificated,to elucidate methods of cells culture in vitro and its cryopereservation,resuscitation and the value of their application. Methods The fibrobiasts from eight cases of keloid and eight cases of normal skin were cultured respectively in vitro.Cells were subcultured.Observing cell morpHology of normal skin and keloid fibroblasts, growth curve was used to evaluate the proliferation of cells.Cells in logarithmic growth pHase were placed in refrigerator for gradient cooling and then were preserved in liquid Nitrogen（-196℃） for 2-6 months, the viability of cells was determined by trypan-blue dye,cell morpHology were also observed. Results The results indicated that the fibroblasts isolated from normal skin and keloid exhibited similar morpHology and growth rates, but fibroblasts of keloid were arranged foul-up,overlap,the polarity were disappeared.The primary cells of fibroblasts from keloid grew faster.The frozen ceils were resuscitated successfully after 2-6 months,the resuscitated rates were more than 80%,and there were no changed of its morpHology. Conclusion The fibroblasts isolated from normal skin and keloid exhibited similar morpHology and growth rates.In vitro,we established normal skin and keloid fibroblasts cells culturing and its cryopereservation, resuscitation successfully.
出处
《中国美容医学》
CAS
2005年第4期394-396,i0002,共4页
Chinese Journal of Aesthetic Medicine
基金
国家自然科学基金资助项目(30271348)
山东省卫生厅课题资助项目(1999CAICJB5)
关键词
瘢痕疙瘩
成纤维细胞
细胞培养
冻存
复苏
keloid
fibroblast
cell culture
refrigerated storage
revivification
