Yan W Q, Tong M H, Yu L, Yu T, Hou L Z, Yang T S, Gao G, Zhang J Y
Institute of Preclinical Sciences, Norman Betheune University Medical Sciences, Changchun.
Shi Yan Sheng Wu Xue Bao. 1994 Jun;27(2):193-203.
Growth-plate cartilage is organized into four cellular zones containing resting, proliferating, maturing, and hypertrophic cells. Rabbit chondrocytes were isolated from growth-plate costal cartilage of 4-week-old New Zealand rabbits, the cells (15 x 10(4)) were suspended in 1 ml of Iscove's modified Dulbecco's medium (IMDM) with 10% fetal bovine serum, 50 micrograms ascorbic acid, and 60 micrograms kanamycin (medium A), then transferred to a 15 ml of plastic centrifuge tube, and centrifuged at 1500 rpm for 5 min. The cell pellet was incubated at 37 degrees C under 5% CO2 in air. The cultures reorganized into growth plate-like tissue which could be seen 7-14 days after cell seeding. This growth-plate, histologically, was organized longitudinally into cellular columns and horizontally into four cellular zones containing resting, proliferating, maturing and hypertrophic cells. The hypertrophic cells in the upper were large in size and round or oval in shape, the proliferating and the mature chondrocytes in the lower were small in size and spherical or elongated in shape. These chondrocytes were surrounded by an extensive matrix. Biochemically, DNA content of cultures began to rise on the 2nd day after cell seeding and reached a plateau after 10 days later. The uronic acid content increased from day 4 and reached the maximum on day 15. In contrast in the early culture, alkaline phosphatase activity was extremely low, it began to rise on day 9 and was the highest on day 20. The sequential increase of DNA, uronic acid and alkaline phosphatase contents was analogous to the in vivo changes of growth-plate chondrocytes.
生长板软骨组织分为四个细胞区,包含静止细胞、增殖细胞、成熟细胞和肥大细胞。从4周龄新西兰兔的生长板肋软骨中分离出兔软骨细胞,将细胞(15×10⁴)悬浮于1ml含有10%胎牛血清、50μg抗坏血酸和60μg卡那霉素的伊思柯夫改良杜尔贝科培养基(IMDM)中(培养基A),然后转移至15ml塑料离心管中,以1500rpm离心5分钟。细胞沉淀在37℃、5%二氧化碳的空气中孵育。细胞接种后7 - 14天可见培养物重组为生长板样组织。该生长板在组织学上纵向组织成细胞柱,横向组织成四个细胞区,包含静止细胞、增殖细胞、成熟细胞和肥大细胞。上部的肥大细胞体积大,呈圆形或椭圆形,下部的增殖细胞和成熟软骨细胞体积小,呈球形或细长形。这些软骨细胞被广泛的基质包围。在生物化学方面,培养物的DNA含量在细胞接种后第2天开始上升,并在10天后达到平台期。糖醛酸含量从第4天开始增加,并在第15天达到最大值。相比之下,在培养早期,碱性磷酸酶活性极低,在第9天开始上升,并在第20天达到最高。DNA、糖醛酸和碱性磷酸酶含量的顺序增加类似于生长板软骨细胞的体内变化。