ANCHORAGE-INDEPENDENT GROWTH OF NORMAL HUMAN-FIBROBLASTS
ANCHORAGE-INDEPENDENT GROWTH OF NORMAL HUMAN-FIBROBLASTS
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DOI：
10.1073/pnas.78.5.3053
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发表时间：
1981-01-01
期刊：
影响因子：
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通讯作者：
STANBRIDGE, EJ
中科院分区：
文献类型：
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作者：
PEEHL, DM;STANBRIDGE, EJ
正常人成纤维细胞，被认为是完全依赖于锚定的增殖，已在甲基纤维素培养基中生长。悬浮液中生长所需的最重要因素似乎是使用高水平的血清和氢化可的松。新生儿包皮[HF 1031、HF 0814、HF 1010和HF 0813]或胎儿肺成纤维细胞（GM 2291）在3周后形成直径为0.5 mm的集落，集落形成效率高达70%。在锚定非依赖性生长的标准测定中不形成集落的小鼠3 T3细胞也在这些条件下生长。接种低至100个细胞/60 mm培养皿后的集落形成结果，并且中期细胞已用荧光DNA染色剂可视化，表明集落形成是由于分裂而不是聚集。从悬浮液中回收并以单层生长的成纤维细胞保留二倍体核型和正常形状，在注射到裸鼠中时不形成肿瘤，并且变得衰老。因此，正常人成纤维细胞明显具有体外锚定非依赖性生长的特性，并且可以在适当的条件下表达。[Also在本研究中使用的是：人宫颈癌HeLa D98 OR细胞;非致瘤性成纤维细胞HeLa D98 OR杂合HSH 5细胞和致瘤性SH 5分离体SH 5 T细胞]。
Normal human fibroblasts, considered to be entirely anchorage dependent for proliferation, have been grown in methylcellulose medium. The most important factor required for growth in suspension appears to be the use of high levels of serum and hydrocortisone. Newborn foreskin [HF1031, HF0814, HF1010 and HF0813] or fetal lung fibroblasts (GM2291] form colonies as large as 0.5 mm in diameter after 3 wk, with a colony-forming efficiency as high as 70%. Mouse 3T3 cells that do not form colonies in standard assays for anchorage-independent growth also grow under these conditions. Colony formation results after inoculation of as few as 100 cells/60 mm dish, and metaphase cells have been visualized with a fluorescent DNA stain, showing that colony formation is due to division rather than aggregation. Fibroblasts recovered from suspension and grown as monolayers retain a diploid karyotype and normal shape, do not form tumors upon injection into nude mice, and become senescent. Thus, the trait of anchorage-independent growth in vitro is clearly possessed by normal human fibroblasts and can be expressed under the proper conditions. [Also used in this study were: human cervical carcinoma HeLa D98OR cells; nontumorisenic fibroblast HeLa D98OR hybrid HSH5 cell and the tumorogenic SH5 segregant SH5T cell.].