Peehl D M, Stanbridge E J
Proc Natl Acad Sci U S A. 1981 May;78(5):3053-7. doi: 10.1073/pnas.78.5.3053.
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 or fetal lung fibroblasts 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 per 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.
正常人类成纤维细胞被认为完全依赖贴壁才能增殖，但它们已在甲基纤维素培养基中生长。悬浮生长所需的最重要因素似乎是使用高浓度血清和氢化可的松。新生儿包皮或胎儿肺成纤维细胞在3周后可形成直径达0.5毫米的集落，集落形成效率高达70%。在非贴壁依赖性生长的标准检测中不形成集落的小鼠3T3细胞在这些条件下也能生长。每60毫米培养皿接种低至100个细胞后即可形成集落，中期细胞可用荧光DNA染色观察到，这表明集落形成是由于细胞分裂而非聚集。从悬浮培养物中回收并作为单层培养的成纤维细胞保留二倍体核型和正常形态，注射到裸鼠体内不会形成肿瘤，且会衰老。因此，正常人类成纤维细胞显然具有体外非贴壁依赖性生长的特性，并且在适当条件下可以表达出来。
