| 朱静 张汝芝 程赛 刘小明 金慧玲.体外人表皮细胞对胰蛋白酶耐受的探讨[J].,2015,15(7):1245-1248 |
| 体外人表皮细胞对胰蛋白酶耐受的探讨 |
| The Tolerance of Human Epidermal Cells to Trypsinization in Vitro |
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| DOI： |
| 中文关键词: 人表皮细胞 耐受性 胰酶消化作用 |
| 英文关键词: Human epidermal cells Tolerance Trypsinization |
| 基金项目:国家自然科学基金项目 (811 71516)； 2015 年中华医学会 - 欧莱雅中国人健康毛发研究项目 |
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| 中文摘要: |
| 目 的： 观察人表皮细胞对胰酶消化的耐受能力 ， 通过不同 细胞恢复贴壁时间 不同 来探索分离 和纯化表皮细胞的新方法， 并
探讨胰酶耐受细胞的干性表达，及其与 muse 细胞的可能相关性。 方法： 中性蛋白酶及胰酶消化获取表皮细胞，用 0.25%的胰酶悬
浮表皮细胞， 以 3.0× 105/mL 的细胞密度种植于 12 孔板， 每间 隔半小时或 1 小时终止胰酶 1 孔， 其中胰酶作用 时间 最长达 46 小
时。 记录不同 细胞贴壁时间、生长状态， 并在接种后第 7 天，对贴壁细胞进行 Nestin、 Sox10 抗体免疫细胞化学染色。 结果： 随着胰
酶作用 时间的延长， 贴壁细胞数目递减， 细胞贴壁所用 时间 也延长。 所有孔中最早出 现的贴壁细胞为 树突状细胞， 这些细胞开始
生长缓慢， 大约 4 天后生长迅速， 10 天后部分孔出现鱼群样细胞团。 部分孔经 Nestin、 SOX10 抗体的免疫细胞化学染色结果均为
阳性，其中以 Nestin 抗体较明显。 结论： 人表皮细胞对胰酶消化的耐受能达 46 小时， 从形态学观察判断， 黑素细胞贴壁早于角 质
形成细胞，大多 数贴壁后细胞增殖力强， 干细胞表面标记显示部分阳性。 |
| 英文摘要: |
| Objective:To observe the tolerance of human epidermal cells to trypsinization in vitro. Search for new method of separating and purifying epidermal cells according to the different adherent time, and investigate the tolerant cells express the markers of
stem cells, and the correlation with muse cells.Methods:The single epidermal cells were obtained by separating the epidermis from a
foreskin, then liberating single cells from the epidermis. In these procedures, the dispase II and trypsin were used respectively. The obtained epidermal cells were resuspended in 0.25% trypsin solution and then seeded in 12-well dishes in density of 3.0 × 105 cells /ml.
Each well was neutralized by bovine serum at different time points, with an interval after every half an hour or one hour, up to 46 h. To
record the time of different cell attachment, and observe the growth state of cells. The adherent cells at 7th day after seeding accepted immunocytochemical staining with the antibodies of Nestin, Sox10, respectively.Results:With prolonged trypsinization, the adherent cells
decreased, the time to attach to wells prolonged. The earliest adherent cells in all wells were dendritic cells, which proliferated slowly at
first, then rapidly after 4 days. At the tenth day, some cells arranged in patterns resembling schools of fish. Some cells showed positive
stainings with antibodies of nestin, sox10, respectively.Conclusion:Human epidermal cells could be tolerant to trypsogen for 46 hours.
According to the shape of cells, the earliest adherent cells were melanocytes with strong proliferation ability, other than keratinocytes.
Some cells showed positive markers of stem cells. |
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