Lamar C H, Eller L L, Turek J J
Am J Vet Res. 1987 Mar;48(3):515-8.
Dispersed cell cultures were established from the articular cartilage of the proximal portion of the humerus of young pigs. Articular and epiphyseal portions of the cartilage were separated, minced, and enzymatically dispersed, using bacterial collagenase. Morphologically, 2 cell types were observed, using phase-contrast microscopy. Smaller polygonal cells (32.5 +/- 3.5 microns diameter) containing cytoplasmic granules were found in both areas of the cartilage. In cultures from the articular region, cells grew as monolayer cultures and initially did not demonstrate contact inhibition. In cultures from the epiphyseal region, cells grew in a multilayered manner in a colonial arrangement with cells being released from the center of the colony into the culture medium. Small granular particles (0.03 to 0.08 micron diameter) were secreted by cells in both culture systems. Particle secretion was greater in epiphyseal cultures than in articular cultures with the rate decreasing as confluency was approached. These particles stained positively for lipid and alkaline phosphatase. Acridine orange was also incorporated into the granules. The 2nd cell type, a stellate-shaped cell (60 +/- 7.6 micron diameter), was found mainly surrounding the outside of colonial areas in epiphyseal cultures. These cells did not secrete small granular particles and stained positive for factor VIII. Evaluation of cultures by scanning and transmission electron microscopy further supported the presence of 2 cell types. With scanning electron microscopy, the smaller polygonal cell was characterized by varying sizes of blebs (0.03 to 0.1 micron diameter) associated with the cell membrane and small cytoplasmic processes projecting from the cell's surface.(ABSTRACT TRUNCATED AT 250 WORDS)
从幼年猪肱骨近端的关节软骨建立了分散细胞培养物。使用细菌胶原酶将软骨的关节部分和骨骺部分分离、切碎并酶解分散。通过相差显微镜在形态学上观察到两种细胞类型。在软骨的两个区域都发现了较小的多边形细胞(直径32.5±3.5微米),含有细胞质颗粒。在关节区域的培养物中,细胞以单层培养物生长,最初未表现出接触抑制。在骨骺区域的培养物中,细胞以多层方式呈集落排列生长,细胞从集落中心释放到培养基中。在两种培养系统中细胞都分泌小颗粒(直径0.03至0.08微米)。骨骺培养物中的颗粒分泌比关节培养物中的更多,随着汇合度接近,分泌速率降低。这些颗粒对脂质和碱性磷酸酶染色呈阳性。吖啶橙也掺入颗粒中。第二种细胞类型是星状细胞(直径60±7.6微米),主要发现在骨骺培养物中集落区域的外部周围。这些细胞不分泌小颗粒,并且对因子VIII染色呈阳性。通过扫描电子显微镜和透射电子显微镜对培养物的评估进一步支持了两种细胞类型的存在。通过扫描电子显微镜,较小的多边形细胞的特征是与细胞膜相关的大小不同的泡(直径0.03至0.1微米)以及从细胞表面伸出的小细胞质突起。(摘要截短于250字)