中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2009年
1期
49-51
,共3页
赵志国%丁克云%金城%陆洪光%尹雪锋
趙誌國%丁剋雲%金城%陸洪光%尹雪鋒
조지국%정극운%금성%륙홍광%윤설봉
黑素细胞%成黑素细胞%细胞培养技术%显微镜检查,电子
黑素細胞%成黑素細胞%細胞培養技術%顯微鏡檢查,電子
흑소세포%성흑소세포%세포배양기술%현미경검사,전자
Melanocytes%Melanoblast%Cell culture techniques%Microscopy,electron
目的 探讨人包皮组织来源成黑素细胞的培养条件.方法 包皮取自小儿,采用分离酶和胰酶两步消化法获得表皮细胞悬液,用实验室改进的成黑素细胞培养基培养结合差速黏附法分选成黑素细胞.对培养的成黑素细胞生长情况进行对比观察,并用Dopa染色、TRP2抗体染色、S-100抗体染色以及电镜观察等方法对培养的成黑素细胞进行鉴定.结果 培养的人黑素细胞呈火柴棒状,散在排列,_双极对称,胞体瘦小,细胞核折光性强,TRP2抗体染色、S-100抗体染色、Dopa染色均阳性,电镜下可见胞内含有Ⅲ一Ⅳ期的成熟黑素颗粒.与黑素细胞相比,培养的人成黑素细胞胞浆丰富,体积较大,呈双极或不规则形,克隆样生长,胞内无Ⅲ～Ⅳ期的成熟黑素颗粒,只含有Ⅰ期不成熟黑素颗粒,TRF2抗体染色阳性、S-100抗体染色、Dopa染色阴性.结论 改良的成黑素细胞培养基成功培养出包皮组织来源的成黑素细胞,为表皮色素形成机制的研究打下基础.
目的 探討人包皮組織來源成黑素細胞的培養條件.方法 包皮取自小兒,採用分離酶和胰酶兩步消化法穫得錶皮細胞懸液,用實驗室改進的成黑素細胞培養基培養結閤差速黏附法分選成黑素細胞.對培養的成黑素細胞生長情況進行對比觀察,併用Dopa染色、TRP2抗體染色、S-100抗體染色以及電鏡觀察等方法對培養的成黑素細胞進行鑒定.結果 培養的人黑素細胞呈火柴棒狀,散在排列,_雙極對稱,胞體瘦小,細胞覈摺光性彊,TRP2抗體染色、S-100抗體染色、Dopa染色均暘性,電鏡下可見胞內含有Ⅲ一Ⅳ期的成熟黑素顆粒.與黑素細胞相比,培養的人成黑素細胞胞漿豐富,體積較大,呈雙極或不規則形,剋隆樣生長,胞內無Ⅲ～Ⅳ期的成熟黑素顆粒,隻含有Ⅰ期不成熟黑素顆粒,TRF2抗體染色暘性、S-100抗體染色、Dopa染色陰性.結論 改良的成黑素細胞培養基成功培養齣包皮組織來源的成黑素細胞,為錶皮色素形成機製的研究打下基礎.
목적 탐토인포피조직래원성흑소세포적배양조건.방법 포피취자소인,채용분리매화이매량보소화법획득표피세포현액,용실험실개진적성흑소세포배양기배양결합차속점부법분선성흑소세포.대배양적성흑소세포생장정황진행대비관찰,병용Dopa염색、TRP2항체염색、S-100항체염색이급전경관찰등방법대배양적성흑소세포진행감정.결과 배양적인흑소세포정화시봉상,산재배렬,_쌍겁대칭,포체수소,세포핵절광성강,TRP2항체염색、S-100항체염색、Dopa염색균양성,전경하가견포내함유Ⅲ일Ⅳ기적성숙흑소과립.여흑소세포상비,배양적인성흑소세포포장봉부,체적교대,정쌍겁혹불규칙형,극륭양생장,포내무Ⅲ～Ⅳ기적성숙흑소과립,지함유Ⅰ기불성숙흑소과립,TRF2항체염색양성、S-100항체염색、Dopa염색음성.결론 개량적성흑소세포배양기성공배양출포피조직래원적성흑소세포,위표피색소형성궤제적연구타하기출.
Objective To study the in vitro culture condition for melanoblasts from human foreskin tissue. Methods The skin tissue taken from foreskin of children was treated with 0.5% dispase Ⅱ to separate epidermis from dermis, then with trypsin to obtain single cell suspension, which was cultured in modified medium for melanoblasts, i.e., MCDB254 medium supplied with several cell growth factors. Finally, melanoblasts were obtained based on the difference of adhesion speed. The morphology and proliferation of cultured melanoblasts were observed under a light microscope. DOPA staining, immunostaining with anti- S-100 and -tyrosinase related protein 2 (TRP2) antibodies, and transmission electron microscopy were per- formed to identify the cultured melanoblasts. Results The cultured human melanocytes displayed a match-like shape, scattered arrangement, syrmnetric double poles, slim cell body, highly refractive nuclei; meanwhile, the melanoblasts exhibited plentiful cytoplasm, large volume, bipolar or irregular shape and clonal growth. Additionally, the melanocytes were positive for TRP2, S-100 and Dopa staining, while the melanoblasts were positive only for TRP2. Electron microscopy revealed the presence of mature melanin granules (stage Ⅲ-Ⅳ ) in melanocytes but immature melanin granules (stage Ⅰ ) in melanoblasts. Conclu- sion Stable pure culture of melanoblasts has been realized with the reformed medium, which may lay a foundation for the investigation into the mechanism of epidermal pigmentation.
