Zeydel M, Puglia K, Eghbali M, Fant J, Seifter S, Blumenfeld O O
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461.
Cell Tissue Res. 1991 Aug;265(2):353-9. doi: 10.1007/BF00398083.
In the heart of the adult rat, fibroblasts are mainly responsible for the synthesis and deposition of the collagenous matrix. Because these cells in vitro may serve as an important model system for studies of collagen metabolism in heart tissue, we have cultured and characterized rat-heart fibroblasts from young adult and old animals. Conditions included use of media of different compositions with and without addition of ascorbate. Cells used were either cultured directly from fresh tissues or thawed previously frozen cells. Cultured cells were studied with respect to growth properties, morphology and ultrastructure and patterns of collagen. Heart fibroblasts generally resembled fibroblasts cultured from other tissues, but were more like skeletal muscle fibroblasts in that they deposited, in addition to type I collagen, type IV collagen and laminin. The fibroblasts showed a typical appearance in phase-contrast microscopy and electron microscopy. In the case of cells grown with added ascorbate, aligned collagen fibrils in the extracellular matrix showed a periodicity typical of type I collagen. The deposition of type I collagen occurred only in medium supplemented with ascorbate, and in that circumstance increased as a function of time past confluence; this was independent of the age of the animal from which the cells were obtained or of other changes of medium composition studied. Immunofluorescence studies with specific antibodies revealed that the cells deposited types I and IV collagens, laminin and fibronectin. In contrast to the case of type I collagen, the deposition of type IV collagen occurred in cells grown either with or without ascorbate.(ABSTRACT TRUNCATED AT 250 WORDS)
在成年大鼠心脏中,成纤维细胞主要负责胶原基质的合成与沉积。由于这些体外培养的细胞可作为研究心脏组织胶原代谢的重要模型系统,我们已从年轻成年和老年动物中培养并鉴定了大鼠心脏成纤维细胞。培养条件包括使用添加或不添加抗坏血酸的不同成分培养基。所用细胞要么直接从新鲜组织中培养,要么解冻先前冷冻的细胞。对培养的细胞进行了生长特性、形态和超微结构以及胶原模式的研究。心脏成纤维细胞通常与从其他组织培养的成纤维细胞相似,但更类似于骨骼肌成纤维细胞,因为它们除了沉积I型胶原外,还沉积IV型胶原和层粘连蛋白。成纤维细胞在相差显微镜和电子显微镜下呈现出典型外观。对于添加抗坏血酸培养的细胞,细胞外基质中排列的胶原纤维呈现出I型胶原特有的周期性。I型胶原的沉积仅发生在添加抗坏血酸的培养基中,且在这种情况下,随着汇合后时间的延长而增加;这与获取细胞的动物年龄或所研究的其他培养基成分变化无关。用特异性抗体进行的免疫荧光研究表明,这些细胞沉积I型和IV型胶原、层粘连蛋白和纤连蛋白。与I型胶原不同,IV型胶原的沉积在添加或不添加抗坏血酸培养的细胞中均会发生。(摘要截选至250词)