Rønnov-Jessen L, Van Deurs B, Nielsen M, Petersen O W
Department of Anatomy, Panum Institute, University of Copenhagen, Denmark.
In Vitro Cell Dev Biol. 1992 Apr;28A(4):273-83. doi: 10.1007/BF02634244.
Myofibroblasts from human breast carcinomas were identified and experimentally generated in culture, and a possible function was examined. The frequency of alpha-smooth muscle actin immunoreactive cells was evaluated as a measure of myofibroblast differentiation in primary culture. Few or no alpha-smooth muscle actin-positive stromal cells (6.1 +/- 8.4%) were identified in primary cultures from normal breast tissue (n = 9). In contrast, high frequencies (68.8 +/- 15.1%) were observed in primary cultures from carcinomas (n = 19). The frequencies of myofibroblasts in primary cultures were almost identical to those obtained in the corresponding cryostat sections (69.1 vs. 68.8%). A possible precursor cell to the myofibroblast was looked for among typical fibroblasts and vascular smooth muscle cells. Purified blood vessels containing both fibroblasts and vascular smooth muscle cells were embedded in collagen gel and incubated with medium conditioned by breast epithelial cells. Fibroblasts rather than smooth muscle cells were recruited from the blood vessels. In medium conditioned by carcinoma cell lines or in co-cultures of carcinoma cell lines and purified fibroblasts, alpha-smooth muscle actin and the typical myofibroblast phenotype were induced in otherwise alpha-smooth muscle actin-negative fibroblasts. The effect of myofibroblasts on cellular movement--essential to neoplastic cells--was analyzed. Spontaneous motility of tumor cells (MCF-7) was entirely suppressed in a collagen gel assay. Under these conditions tumor cell motility was selectively mediated by direct cell-to-cell interaction between tumor cells and myofibroblasts. Under chemically defined conditions, interaction was dependent on the presence of plasminogen. Anti-plasminogen, soybean trypsin inhibitor, and anti-fibronectin partly neutralized the effect of plasminogen. It is concluded that elements of myofibroblast differentiation and function may be studied in culture.
对来自人乳腺癌的肌成纤维细胞进行了鉴定，并在培养中通过实验生成，同时研究了其可能的功能。评估α-平滑肌肌动蛋白免疫反应性细胞的频率，以此作为原代培养中肌成纤维细胞分化的指标。在来自正常乳腺组织的原代培养物（n = 9）中，几乎没有或未发现α-平滑肌肌动蛋白阳性的基质细胞（6.1±8.4%）。相比之下，在来自癌组织的原代培养物（n = 19）中观察到高频率（68.8±15.1%）。原代培养物中肌成纤维细胞的频率与相应冰冻切片中的频率几乎相同（69.1%对68.8%）。在典型的成纤维细胞和血管平滑肌细胞中寻找肌成纤维细胞可能的前体细胞。将含有成纤维细胞和血管平滑肌细胞的纯化血管包埋在胶原凝胶中，并用乳腺上皮细胞条件培养基进行孵育。从血管中募集的是成纤维细胞而非平滑肌细胞。在癌细胞系条件培养基中或在癌细胞系与纯化成纤维细胞的共培养物中，原本α-平滑肌肌动蛋白阴性的成纤维细胞可被诱导表达α-平滑肌肌动蛋白和典型的肌成纤维细胞表型。分析了肌成纤维细胞对细胞运动（这对肿瘤细胞至关重要）的影响。在胶原凝胶试验中，肿瘤细胞（MCF-7）的自发运动性被完全抑制。在这些条件下，肿瘤细胞的运动性由肿瘤细胞与肌成纤维细胞之间的直接细胞间相互作用选择性介导。在化学限定条件下，这种相互作用依赖于纤溶酶原的存在。抗纤溶酶原、大豆胰蛋白酶抑制剂和抗纤连蛋白可部分中和纤溶酶原的作用。得出的结论是，可以在培养中研究肌成纤维细胞分化和功能的相关因素。
