白细胞介素-23对人表皮黑素细胞生物活性的影响

目的观察白细胞介素(IL)-23对黑素细胞增殖、凋亡及酪氨酸酶活性的影响,为白癜风发病机制的研究奠定基础。方法建立模拟正常生理状态的黑素细胞体外培养体系,用不同浓度IL-23作用于黑素细胞后,采用CCK-8法检测不同浓度IL-23对黑素细胞增殖活性的影响;流式细胞仪检测其对黑素细胞凋亡的影响;L-DOPA法检测其对黑素细胞酪氨酸酶活性的影响。结果 IL-23随着浓度的增加抑制黑素细胞的增殖,与空白对照组相比差异有统计学意义(P0.05);用浓度为100 ng/m L的IL-23培养黑素细胞,48 h后细胞凋亡率分别为23.0%,空白对照组为5.1%;100 ng/m L和200 ng/m L IL-23组抑制酪氨酸酶的活性,与空白对照组相比差异有统计学意义(P0.05)。结论一定浓度的IL-23对体外培养的黑素细胞起抑制增殖,并促进细胞凋亡,抑制酪氨酸酶活性的作用,其可能参与白癜风的发生发展,但其作用机制还需进一步实验和探讨。     

他卡西醇对人表皮黑素细胞增殖、黏附、迁移及c-kit表达的影响

【摘要】 目的 探讨他卡西醇对体外培养的人表皮黑素细胞增殖、黏附、迁移及c-kit mRNA相对表达的影响。 方法 用不同浓度的他卡西醇干预体外培养的人表皮黑素细胞,采用四唑氮氢氧化物（XTT）法分别检测培养24、48、72 h后黑素细胞的增殖活性;用纤维连接蛋白（FN）包被细胞培养板检测培养72 h后黑素细胞的黏附率;用Transwell微孔膜法检测培养24 h后黑素细胞在FN上的迁移情况;逆转录-聚合酶链反应（RT-PCR）检测培养72 h后黑素细胞c-kit mRNA相对表达量。 采用重复测量方差分析及完全随机设计方差分析进行统计学检验。 结果 重复测量方差分析结果显示,10-10、10-9、10-8、10-7、10-6 mol/L他卡西醇促进黑素细胞增殖作用的差异有统计学意义（F = 9.47,P < 0.01）,上述浓度他卡西醇在培养24、48、72 h后促进黑素细胞增殖作用的差异有统计学意义（F = 14.44,P < 0.01）,药物浓度和培养时间之间存在交互作用（F = 2.47,P < 0.01）,其中10-8 mol/L他卡西醇与黑素细胞共同培养72 h黑素细胞增殖活性最高。10-8 ～ 10-7 mol/L他卡西醇可显著促进黑素细胞在FN上的黏附（均P < 0.01）;10-9 ～ 10-8 mol/L他卡西醇在培养24 h能显著促进黑素细胞迁移（均P < 0.01）;RT-PCR显示10-9 ～ 10-7 mol/L他卡西醇在培养72 h均能显著增加黑素细胞c-kit mRNA的相对表达量（均P < 0.01）。 结论 他卡西醇可促进培养的人表皮黑素细胞增殖及在FN上的黏附、迁移作用,并可上调黑素细胞c-kit mRNA的表达。 【关键词】 他卡西醇; 黑素细胞; 细胞增殖; 细胞运动; 原癌基因蛋白质c-kit     

皮肤科学基础

20062382碱性成纤维细胞生长因子和α-黑素细胞刺激素对体外培养黑素细胞黏附和迁移的影响/张宪旗(浙江大学医学院二附院皮肤科),冯捷,牟宽厚…∥浙江大学学报.-2006,35(2).-161~164从正常人包皮分离、纯培养黑素细胞,分别用碱性成纤维细胞生长因子(bFGF)和α-黑素细胞刺激素(α-MSH)处理培养黑素细胞;采用纤维连接素包裹的培养板观测黑素细胞黏附,用Transwell微孔膜法研究黑素细胞迁移。结果显示bFGF和α-MSH均可促进黑素细胞黏附于培养板,bFGF可诱导黑素细胞迁移并呈剂量依赖性,α-MSH对黑素细胞迁移无明显影响。提示bF-GF和α-…     

白细胞介素-22对人表皮黑素细胞生物活性的影响

目的观察白细胞介素(IL)-22对黑素细胞生物活性的影响,包括黑素细胞的增生、黑素细胞酪氨酸酶活性和黑素细胞凋亡,从而研究色素障碍性皮肤病的发病机制,为色素性疾病的治疗提供新的思路。方法建立模拟正常生理状态的黑素细胞体外培养体系,用不同浓度IL-22作用于黑素细胞,采用CCK-8法检测不同浓度IL-22对黑素细胞增生活性的影响;L-DOPA法检测其对黑素细胞酪氨酸酶活性的影响;流式细胞仪检测其对黑素细胞凋亡的影响。结果 1 IL-22随着浓度的增加呈明显的时间依赖性抑制黑素细胞的增生,与空白对照组相比差异有统计学意义(P0.05)。2用浓度为100 ng/ml的IL-22培养黑素细胞,48 h后细胞凋亡率为23.0%,空白对照组为5.1%。结论 IL-22随着浓度的增加明显抑制黑素细胞的增生,呈时间依赖性,当培养时间为72 h时,所有浓度均抑制黑素细胞的增生,黑素细胞酪氨酸酶活性也随着IL-22浓度的增加而降低,黑素细胞凋亡率较空白对照组明显升高,均提示IL-22可引起黑素细胞的损伤。     

白介素-10,白介素-17对人表皮黑素细胞生物活性的影响

目的观察不同浓度白介素-10,白介素-17对黑素细胞生物活性的影响。方法建立模拟正常生理状态的黑素细胞体外培养体系,用不同浓度白介素-10,白介素-17作用于黑素细胞后,采用CCK-8法测定白介素-10,白介素-17对黑素细胞增殖活性的影响、L-DOPA法测定对黑素细胞酪氨酸酶活性的影响、流式细胞仪检测黑素细胞凋亡率。结果 1 50μg/ml IL-10组对黑素细胞的增殖率随着培养时间的延长而升高,当培养48h和72h后与空白对照组相比有极显著的统计学差异(P0.01)。但随着IL-10浓度的升高,其增殖率随着培养时间的延长反而有下降趋势,200μg/ml IL-10组培养黑素细胞48h和72h后与空白对照组相比有统计学差异(P0.05);2 IL-17随着培养时间的延长呈剂量依赖性抑制黑素细胞的增殖,50μg/ml IL-17组培养黑素细胞48h和72h后与空白对照组相比有统计学差异(P0.05);100μg/ml IL-17组和200μg/ml IL-17组培养黑素细胞48h和72h后与空白对照组相比有极显著的统计学差异(P0.01);3在培养48h后,50μg/ml IL-10组黑素细胞酪氨酸酶活性升高,较空白对照组有统计学差异(P0.05)。50μg/ml、100μg/ml、200μg/ml IL-17组抑制黑素细胞酪氨酸酶的活性,与空白对照组相比有统计学差异(P0.05);4用浓度为100μg/ml的IL-10,IL-17培养黑素细胞,48小时后细胞凋亡率分别为5.2%、32.1%,空白对照组为5.1%。结论 1适当浓度的IL-10可促进黑素细胞的增殖,提高酪氨酸酶的活性;2 IL-17对黑素细胞有损伤作用,但其作用机制还需进一步的试验和探讨。     

白细胞介素-17对人黑素细胞合成黑素功能的影响

目的研究细胞因子白细胞介素(IL)-17对体外培养的人黑素细胞合成黑素功能的影响。方法不同浓度的细胞因子IL-17A作用于体外培养的人黑素细胞48 h,用L-DOPA法检测黑素细胞的酪氨酸酶活性,Na OH溶解法检测黑素合成量。荧光定量PCR检测高浓度IL-17A对黑素细胞酪氨酸酶、酪氨酸酶相关蛋白-1和MITF等基因m RNA表达的影响,Annexin V/PI双色标记法结合流式细胞术检测高浓度IL-17A诱导黑素细胞凋亡的比例。结果不同浓度的细胞因子IL-17A对黑素细胞酪氨酸酶活性及黑素的合成具有一定的抑制作用。高浓度的IL-17A可抑制酪氨酸酶、酪氨酸酶相关蛋白-1和MITF基因m RNA的表达,但无明显诱导黑素细胞凋亡的作用。结论 IL-17可通过抑制黑素合成关键基因m RNA的表达而在一定程度上抑制黑素细胞合成黑素,可能参与了白癜风黑素细胞的损伤。但因其无直接诱导黑素细胞凋亡的作用,推测并非是导致白癜风黑素细胞丢失的主要原因。     

芍药苷对人黑素细胞生物活性及迁移的影响

目的:确定芍药苷对人黑素细胞生物活性及迁移的影响。方法:体外培养人表皮黑素细胞,分别使用MTT法、L-DOPA法、NaOH法、Transwell小室法检测不同浓度的芍药苷对黑素细胞的增殖活性、酪氨酸酶活性、黑素合成以及黑素细胞迁移的影响。结果:芍药苷能显著促进黑素细胞增殖和上调酪氨酸酶活性,且10μ/mL是产生这两方面作用的最佳浓度;芍药苷也能促进黑素的合成,50μg/mL时作用最显著。芍药苷在20μg/mL时显著促进黑素细胞迁移。各组与空白对照组比较差异均有显著性(P0.05)。结论:芍药苷单体可能是白芍中促进黑素细胞迁移的主要活性成分。     

复方中药对黑素细胞迁移和黏附的影响

目的:确定复方中药乙醇提取物对体外培养黑素细胞黏附、迁移的影响.方法:复方中药用70%乙醇提取,正常人黑素细胞来自包皮环切术切取的包皮组织.用包被纤维黏连蛋白的培养板检测黑素细胞黏附,用微孔膜研究黑素细胞迁移.结果:复方中药1号和3号乙醇提取物可促进黑素细胞黏附和迁移(P<0.05),而1、2和3号可以促进黑素细胞迁移(P<0.05).结论:部分复方中药可能通过促进黑素细胞黏附和迁移途径对白癜风起治疗作用.     

土茯苓粗提物对11种黑素细胞株增殖和凋亡的影响

目的:观察土茯苓提取物(rhizoma smilacis glabrae extract,RSGE)对黑素细胞株增殖的抑制作用,并探讨其机制。方法:CCK8法观察RSGE对黑素细胞株及正常人黑素细胞活性影响;流式细胞仪检测RSGE对细胞凋亡及周期的影响;采用反转录(RT)-PCR法检测黑素瘤A2058、HT-144等11种细胞株和正常人黑素细胞中c-kit m RNA表达情况;western blot法检测RSGE对聚腺苷二磷酸核糖聚合酶(PARP)、c-kit等蛋白的影响;real-time PCR法检测RSGE对c-kit基因转录水平的影响。结果:RSGE对c-kit m RNA高表达黑素细胞株A2058增殖的抑制作用强于低表达株HT-144,RSGE处理A2058细胞24 h半抑制浓度(IC50)为(10.55±0.29)g/L;对正常人黑素细胞生长无影响。结论:RSGE通过抑制c-kit基因表达,诱导黑素细胞凋亡,从而发挥抗癌作用。     

内皮素-1和干细胞因子对黑素细胞肌动蛋白的影响

目的 探讨细胞因子内皮素-1(ET-1)和干细胞因子(SCF)对培养的黑素细胞肌动蛋白的作用.方法 采用免疫荧光技术观察黑素细胞在受到ET-1,SCF作用后细胞肌动蛋白actin在细胞内分布情况,间接判断其对细胞行为的影响.结果 黑素细胞在细胞因子的作用下,细胞肌动蛋白呈现聚集性变化,细胞活动功能增强.尤其以ET-1作用更明显(P<0.05),而actin 活动功能的增加直接关系着黑素细胞的粘附和迁移.结论 ET-1和SCF在体外均可促进黑素细胞肌动蛋白actin的聚集,从而间接发挥它们促进黑素细胞粘附和迁移的作用.     

Co-culture of melanocytes with adipose-derived stem cells as a potential substitute for co-culture with keratinocytes

Cell-to-cell interactions between melanocytes and keratinocytes increase the proliferation and migration of melanocytes. In fact, mixed keratinocyte and melanocyte cultures have been used for autologous cell transplantation for treatment of vitiligo. However, this may require taking an amount of skin tissue large enough to leave scars. In this study, the in vitro effect of adipose-derived stem cells (ADSCs) on proliferation, differentiation and migration of melanocytes was compared with that of keratinocytes using immunohistochemistry and a Boyden chamber migration assay. The proliferation and migration of melanocytes was significantly stimulated by co-culture with ADSCs compared with melanocyte monocultures, al-though the effect of ADSCs was less powerful than that of keratinocytes. This may be related to increases in stem cell factor and basic fibroblast growth factor, growth factors for melanocytes, produced by the ADSCs. The ratios of melanocytes stained with antibodies against Trp-2, E-cadherin and N-cadherin were significantly increased by co-culturing with ADSCs compared with co-culturing with keratinocytes as well as melanocyte monocultures. The proportion of less-pigmented melanocytes was also increased and sustained for a longer duration in the presence of ADSCs. Our data show that co-culturing with ADSCs results in increased melanocyte proliferation and migration while reducing differentiation, and could provide a means to treat disorders such as vitiligo.     

壳聚糖膜转载黑素细胞动物实验研究

目的 探讨壳聚糖膜搭载、转运黑素细胞的可行性,探索利用壳聚糖膜改进黑素细胞移植技术。方法 使用电镜、MTT法、NaOH裂解法等技术观察黑素细胞在壳聚糖膜上贴附、生长、合成黑素情况。进行动物移植实验,免疫组化结合激光共聚焦显微镜技术检测壳聚糖膜上搭载的黑素细胞向动物皮肤创面转移情况。结果 扫描电镜和倒置显微镜观察到黑素细胞在壳聚糖膜上分布均匀,贴附良好。利用MTT法测定的增殖曲线显示,壳聚糖膜能够支持黑素细胞正常生长;黑素合成量（A值为0.087 ± 0.027）与培养皿所培养细胞（0.101 ± 0.036）相比差异无统计学意义（P > 0.05）。激光共聚焦显微镜观察裸鼠移植部位,可以见到黑素细胞;在移植部位活检取材,用melan-A单克隆抗体免疫组化染色,结果阳性。结论 壳聚糖膜能够搭载黑素细胞贴附、生长,并支持黑素细胞向动物移植部位转移,同时保持黑素细胞活性。     

Narrow-band ultraviolet-B stimulates proliferation and migration of cultured melanocytes

Narrow-band ultraviolet-B (UVB) radiation is an effective treatment for vitiligo vulgaris. However, the mechanisms of narrow-band UVB in inducing repigmentation of vitiligo lesions are not thoroughly clarified. The purpose of our study was to investigate the effects of narrow-band UVB irradiation on melanocyte proliferation and migration in vitro. Our results showed that the cell counts as well as [3H]thymidine uptake of melanocytes were significantly enhanced by narrow-band UVB-irradiated keratinocyte supernatants. In these supernatants, a significant increase in basic fibroblast growth factor (bFGF) and in endothelin-1 (ET-1) release was observed. bFGF is a natural mitogen for melanocytes, whereas ET-1 can stimulate DNA synthesis in melanocytes. This stimulatory effect of melanocyte proliferation by supernatants derived from narrow-band UVB-irradiated keratinocytes was significantly reduced by a selective endothelin-B (ET-B) receptor antagonist (BQ788), suggesting an essential role of ET-1 on melanocyte proliferation. Our results of time-lapse microphotography revealed a stimulatory effect of narrow-band UVB irradiation on melanocyte migration. Focal adhesion kinase (FAK) plays a pivotal role in cell migration. Phosphorylated FAK (p125(FAK)) expression on melanocyte was enhanced by narrow-band UVB irradiation. In this study, narrow-band UVB irradiation stimulated a significant increase in matrix metalloproteinase-2 (MMP-2) activity in melanocyte supernatants. Narrow-band UVB-irradiation-induced migration of melanocytes was significantly annihilated by the addition of p125(FAK) inhibitor (herbimycin-A) or MMP-2 inhibitor (GM6001). These results suggest that p125(FAK) and MMP-2 activity play important roles in narrow-band UVB-induced migration of melanocytes. Our results provide a theoretical basis for the effectiveness of narrow-band UVB irradiation in treating vitiligo.     

A study of autologous melanocyte transfer in treatment of stable vitiligo

BACKGROUND: Replenishing melanocytes selectively in vitiliginous macules by autologous melanocytes is a promising treatment. With expertise in culturing melanocytes, it has now become possible to treat larger recipient areas with smaller skin samples. AIM: To study the extent of repigmentation after autologous melanocyte transplantation in patients with stable vitiligo. METHODS: The melanocytes were harvested as an autologous melanocyte rich cell suspension from a donor split thickness graft. Melanocyte culture was performed in selected cases where the melanocyte cell count was insufficient to meet the requirement of the recipient area. These cells were then transplanted to the recipient area that had been superficially dermabraded. RESULTS: An excellent response was seen in 52.17% cases with the autologous melanocyte rich cell suspension (AMRCS) technique and in 50% with the melanocyte culture (MC) technique. CONCLUSION: Autologous melanocyte transplantation can be an effective form of surgical treatment in stable but recalcitrant lesions of vitiligo.     

Autologous melanocyte–keratinocyte suspension in the treatment of vitiligo

Background In stable vitiligo, several techniques of autologous transplantation of melanocytes are used. Autologous melanocyte transplantation of non‐cultured melanocytes is one of those techniques with variable reported outcomes. Objective The objective of this study was to evaluate the response to autologous melanocyte–keratinocytes suspension transplantation in cases of stable vitiligo. Methods A total of 25 cases of vitiligo were treated by autologous melanocyte–keratinocytes suspension transplantation. After 6–17 months, patients’ response was evaluated according to the extent of pigmentation (excellent 90–100%, good 50–89%, fair 20–49% and poor response <20%). Results Of the 25 patients treated, 22 continued the follow‐up period. Five (23%) patients showed excellent response, 7 (32%) good, 6 (27%) fair and 4(18%) showed poor response. Conclusion Unlike transplantation of cultured melanocytes, which requires experience in culture technique, autologous melanocyte–keratinocytes suspension transplantation is an easy economic technique, which may be used in resistant areas of stable vitiligo.     

Basic fibroblast growth factor promotes melanocyte migration via increased expression of p125(FAK) on melanocytes

Vitiligo is an acquired pigmentary disorder characterized by depigmentation of skin and hair. Melanocyte migration is an important event in re-pigmentation of vitiligo. We have demonstrated that narrow-band ultraviolet B (UVB) irradiation stimulated cultured keratinocytes to release a significant amount of basic fibroblast growth factor (bFGF). Furthermore, narrow-band UVB enhanced migration of melanocytes via increased expression of phosphorylated focal adhesion kinase (p125(FAK)) on melanocytes. The aim of this study was to investigate the effect of recombinant human bFGF (rhbFGF) on melanocyte migration. The relationship between the expression of p125(FAK) and melanocyte migration induced by rhbFGF was also studied. Our results demonstrated that rhbFGF significantly enhanced migration of melanocytes and p125(FAK) expression on melanocytes. Herbimycin A, a potent p125(FAK) inhibitor, effectively abolished rhbFGF-induced melanocyte migration. The combined results indicated that p125(FAK) plays an important role in the signal transduction pathway of melanocyte migration induced by bFGF.     

Human skin melanocyte migration towards stromal cell‐derived factor‐1α demonstrated by optical real‐time cell mobility assay: modulation of their chemotactic ability by α‐melanocyte‐stimulating hormone

To identify potential regulators of normal human melanocyte behaviour, we have developed an in vitro human melanocyte migration assay, using the optically accessible, real‐time cell motility assay device TAXIScan. Coating of the glass surface with an extracellular matrix that served as scaffolding molecule was essential to demonstrate efficient melanocyte migration. Among several chemokines tested, stromal cell‐derived factor (SDF)‐1α/CXCL12 was the most effective driver of human normal skin melanocytes. Incubation of melanocytes with α‐melanocyte‐stimulating hormone (MSH) before the assay specifically enhanced CXCR4 expression and consequently chemotaxis towards SDF‐1α/CXCL12. These results suggest that α‐MSH acts on melanocytes to produce melanin as well as stimulates the cells to migrate to the site where they work through CXCR4 up‐regulation, which is a new dynamic mode of action of α‐MSH on melanocyte physiology.     

Application of computerized image analysis in pigmentary skin diseases

BACKGROUND: Melanocyte number and the amount of melanin pigment are related to diagnosis and treatment of pigmentary skin diseases. Various histologic methods are used, such as Fontana-Masson stain for melanin pigment or immunohistochemical stain for melanocytes. Recently, computerized image analysis has been applied to many fields to avoid interobserver bias. In this study, we applied a computerized image analysis to assess the melanin content and melanocyte density of human epidermis. METHODS: We evaluated the skin biopsy specimens (paraffin blocks) from normal human skin (33 +/- 6.6, n = 11) and diseased skins; vitiligo (32 +/- 10.0, n = 8), melasma (35 +/- 8.6, n = 11), and lentigo senilis (40 +/- 7.2, n = 11) (mean age +/- SD). Each specimen was stained with Fontana-Masson for melanin pigments and immunohistochemical method for melanocytes. Quantitative analysis of melanin pigment and melanocyte number (density) were investigated through two methods: (1) two dermatologists measured the visual scales; and (2) computerized image analysis was used to measure melanin content indices (MCI). The data were evaluated using one-way ANOVA. RESULTS: The visual scale of the Fontana-Masson stain was the highest for lentigo senilis (3.8 +/- 0.40), followed by melasma (2.6 +/- 0.67), normal skin (1.8 +/- 0.60) and vitiligo (0) (P < 0.05). These findings were consistent with objective measurements made by computerized image analysis. MCI values were 120.3 +/- 20.74 for lentigo senilis, 81.1 +/- 19.27 for melasma, 45.5 +/- 16.92 for normal skin, and 0.3 +/- 0.30 for vitiligo in decreasing order (P < 0.05). MC/1E (melanocyte number per 1 mm epidermis) was about two fold larger in lentigo senilis (18.1 +/- 8.92) than melasma (9.7 +/- 2.40) or normal skin (9.3 +/- 2.67) (P < 0.05). MC/1B (melanocyte number per 1 mm basal layer) was about 1.5 fold higher in lentigo senilis (13.5 +/- 4.17), compared to normal skin (9.0 +/- 3.55) (P < 0.05). Melasma showed increased melanocyte numbers compared to normal skin, but it was not statistically significant (P > 0.05). CONCLUSION: We believe this computerized image analysis could be useful tool for diagnosis and comparison of interval changes in pigmentary diseases like melasma or lentigo senilis by quantifying melanin pigments or melanocytes in skin biopsy specimens.     

甲氧沙林对黑素细胞粘附及基质金属蛋白酶-2的影响

目的 探讨甲氧沙林治疗白癜风的可能机制.方法 正常人黑素细胞来自包皮环切术切取的包皮,用包被纤维黏连蛋白的48孔培养板检测黑素细胞粘附,用带微孔膜的Transwell培养板研究黑素细胞迁移,用RT-PCR方法检测基质金属蛋白酶-2(MMP-2)mRNA表达.结果 50nmol/L和100nmol/L甲氧沙林可促进黑素细胞粘附(P<0.05)和迁移(P<0.05),100nmol/L对黑素细胞促迁移作用最强(P<0.01),而且10nmol/L甲氧沙林即可明显促进MMP-2mRNA表达(P<0.01).在一定浓度下,甲氧沙林促黑素细胞粘附、迁移和MMP-2mRNA表达呈剂量依赖性,在不同浓度组间比较差异均有统计学意义(P<0.01、P<0.05和P<0.01).结论 甲氧沙林通过促进MMP-2的表达来促进黑素细胞粘附和迁移.     

SV40TAg和Cre/loxP系统诱导的正常人可逆性转化黑素细胞在豚鼠体内的存活能力和复色实验

目的 研究SV40TAg基因和Cre/loxP系统在体外诱导正常人黑素细胞的可逆性转化以及转化细胞在豚鼠体内的存活能力和复色效果。方法 用Cre-ERT2病毒上清感染经SV40TAg基因转化的正常人黑素细胞（MCT）,诱导Cre重组酶表达（MCTC）;过氧化氢法建立黄褐色雄性豚鼠白癜风动物模型,按黑素细胞移植方法进行原代黑素细胞和MCTC移植,观察MCTC在豚鼠体内的存活能力及复色效果。结果 MCTC细胞移植实验结果显示,1个月左右移植区即可见色素沉着,连续观察3个月,可见0.5 ～ 1 cm的黑色斑片或褐色斑片形成,移植成功率为82.5%,原代黑素细胞移植成功率为76.7%。病理观察移植区有明显黑素沉积,部分毛囊内也可见黑素沉积。结论 联合应用SV40TAg基因和Cre/loxP系统可以成功地诱导具有良好生物学安全性的人源性可逆性永生化黑素细胞,具有与原代黑素细胞相似的在体存活率,具备在体黑素合成功能,可以达到良好的复色效果。