Absence of tumorigenicity in athymic mice by normal human epidermal keratinocytes after culture in serum-free medium.
Absence of tumorigenicity in athymic mice by normal human epidermal keratinocytes after culture in serum-free medium.
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正常人表皮角质形成细胞在无血清培养基中培养后，无胸腺小鼠不具有致瘤性。
DOI：
10.1016/0304-3835(92)90184-w
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发表时间：
1992
期刊：
影响因子：
9.7
通讯作者：
Hansbrough,JF
中科院分区：
文献类型：
--
作者：
Boyce,ST;Foreman,TJ;Furmanski,P;Hansbrough,JF
正常人表皮角质形成细胞（HK）在无血清培养基中培养的非常快的生长速度（1个群体倍增/天）可用于烧伤治疗中的伤口闭合。然而，在体外快速生长可能存在肿瘤转化的可能性。为了研究这种可能性，在补充有表皮生长因子（EGF，10 ng/ml）、胰岛素（5 μg/ml）、氢化可的松（0.5 μ g/ml）和牛胎盘提取物（BPE，70 μg/ml）的MCDB 153培养基中将HK从原代分离培养到大群体。HK在皮下接种后在无胸腺小鼠中研究其形成肿瘤的能力。从原代培养物或在液氮中储存之前或之后的二次培养物中接种16个单独的HK菌株。转化的细胞系，SCC 13和FL，来源于人上皮癌细胞被用作肿瘤形成的对照。HK培养2 6周未形成肿瘤（0 79），SCC 13培养2 0周形成结节状肿瘤（3 5），FL培养4周形成肿瘤（5 5）。对皮肤原代培养的角质形成细胞接种部位进行组织学检查，未发现HK细胞。相比之下，SCC 13和FL的可触及肿瘤均返回组织培养并继续增殖。这些结果支持了这样的结论，即人表皮角质形成细胞在体外的快速生长速率可以归因于允许的培养条件，而不是肿瘤转化。
The very rapid growth rate (1 population doubling/day) of normal human epidermal keratinocytes (HK) cultured in serum-free medium can be utilized for wound closure in burn treatment. However, rapid growth in vitro may present the possibility of neoplastic transformation. To investigate this possibility, HK were cultured from primary isolation to large populations in MCDB 153 medium supplemented with epidermal growth factor (EGF, 10 ng/ml), insulin (5 μg/ml), hydrocortisone (0.5 μg/ml), and Bovine Pituitary Extract (BPE, 70 μg/ml). HK were studied for their ability to form tumors in athymic mice after subcutaneous inoculation. Sixteen separate HK strains were inoculated from primary cultures, or from secondary cultures either before or after storage in liquid nitrogen. Transformed cell lines, SCC 13 and FL, derived from human epithelial carcinomata were used as controls for tumor formation. HK formed no tumors (0 79) after 26 weeks incubation, SCC 13 formed nodular tumors (3 5) after 20 weeks incubation, and FL formed tumors (5 5) after 4 weeks incubation. HK cells were not found by histological examination of inoculation sites of keratinocyte cultures derived from primary culture from skin. In contrast, palpable tumors from both SCC 13 and FL were returned to tissue culture and continued to proliferate. These results support the conclusion that the rapid growth rate of human epidermal keratinocytes in vitro can be attributed to permissive culture conditions, and not to neoplastic transformation.