SERUM-FREE CULTURE OF TRANSFORMED AND UNTRANSFORMED MOUSE KERATINOCYTES

转化和未转化小鼠角质细胞的无血清培养

• 批准号: 3939672
• 负责人: J FALCO
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3939672
• 关键词: cell bank /registry; cell differentiation; epidermal growth factor; fibroblast growth factor; gene expression; growth media; insulin; keratinocyte; molecular biology; oncogenes; oncogenic virus; skin; tissue /cell culture; transforming virus; viral carcinogenesis

A chemically-defined tissue culture system developed for the BALB/MK epithelial cell line was used to study (1) how the introduction of activated viral oncogenes alters the cell line's growth factor requirements, and (2) what growth factors elicit a mitogenic response in this cell line and how these growth factors interact with each other. In this defined media system, uninfected BALB/MK keratinocytes required only two growth factors for growth--insulin and epidermal growth factor (EGF). Oncovirally infected BALB/MK demonstrated four patterns of growth factor requirements: (1) requirements unaltered from parental line (v-raf); (2) partial escape from EGF requirement (v-mos, v-fms, v-erbB); (3) complete escape from EGF requirement (v-K-ras, v-H-ras); and (4) escape from all growth factor requirements (v-fgr). Three of these viral infectants, v-K-ras, v-fgr, and v-fms, demonstrated release into the medium of EGF-like activity, presumably TGF alpha. No viral infectants produced insulin-like activity. In defined medium mitogenic assays, the following growth factors were found to be BALB/MK mitogens, in descending order of potency: acidic fibroblast growth factor (FGF), basic FGF, EGF, and insulin. Synergism was noted between insulin and EGF or insulin and basic FGF.

一种化学定义的组织培养系统， 本研究以BALB/MK上皮细胞系为研究对象， 引入激活的病毒癌基因改变了细胞系的 生长因子的需求，以及（2）什么生长因子引起 有丝分裂反应以及这些生长因子 相互影响。 在此确定培养基系统中，未感染的BALB/MK角质形成细胞 只需要两种生长因子--胰岛素和 表皮生长因子（EGF） 瘤病毒感染的BALB/MK 证明了生长因子需求的四种模式：（1） 要求与亲本系（v-raf）相同;（2）部分 逃避EGF需求（v-mos，v-fms，v-erbB）;（3） 完全逃避EGF需求（v-K-ras，v-H-ras）;和 (4)逃避所有生长因子需求（V-FGR）。 其中三种病毒感染物v-K-ras v-fgr和v-fms， 证明释放到EGF样活性的介质中， 可能是转化生长因子α 无病毒感染产生胰岛素样 活动 在成分确定的培养基促有丝分裂试验中， 被发现是BALB/MK有丝分裂原，以降序排列， 功效：酸性成纤维细胞生长因子（FGF），碱性FGF，EGF， 和胰岛素。 胰岛素和EGF之间存在协同作用， 胰岛素和碱性FGF。