Jones S J, Ali N N, Boyde A
Anat Embryol (Berl). 1986;174(2):265-75. doi: 10.1007/BF00824342.
Previous studies have shown that osteoclasts obtained from chopped bones resorb surrogate calcified tissue substrata in vitro. These cultures contained all bone and marrow cell types pooled together. We have now parted the marrow from the bone and cultured the cells from the two fractions separately: on both resorbable substrates and on plastic in order to test their longevity in culture and ability to resorb following trypsinisation. Marrow-fraction, bone-fraction or whole bone derived cells were harvested from prehatch chick long bone shafts after removing the periosteum; seeded on sperm whale dentine (SWD) slices or plastic dishes and cultured continuously, or trypsinised and reseeded on to fresh substrata at weekly or half-weekly intervals. Observations were made by light microscopy and SEM. Many multinucleate cells were observed in the marrow fraction immediately after settling, deriving presumably from poorly adherent osteoclasts, next to bone, which had not been resorbing at the time of harvesting. By three days in culture on plastic, multinucleate cells were very large both in terms of plan extent and nuclear number: cell fusion occurred between osteoclasts and between osteoclasts and small, round uninuclear cells. SWD was extensively resorbed. The adherence of the osteoclasts was greater (a) to plastic upon trypsinisation than that of the other cells; and (b) to SWD than to plastic, particularly if the cells were resorbing. Trypsinised cells regained their resorptive capacity after seeding on to new SWD, but only for 1 or 2 treatments. Bone derived cells were similar to the marrow cultures, except for the much higher proportion of other bone cell types. Trypsinisation caused a higher proportional loss of multinucleate cells from both SWD and plastic. Resorption was still occurring at 6 weeks in all cultures. A wide diversity existed in the shapes, depths, plan areas and volumes of the resorption pits.
先前的研究表明，从切碎的骨头中获取的破骨细胞在体外可吸收替代钙化组织基质。这些培养物包含汇集在一起的所有骨和骨髓细胞类型。我们现在已将骨髓与骨分离，并分别培养这两个部分的细胞：在可吸收基质和塑料上，以测试它们在培养中的寿命以及胰蛋白酶消化后吸收的能力。在去除骨膜后，从孵化前的雏鸡长骨干中收获骨髓部分、骨部分或全骨来源的细胞；接种在抹香鲸牙质（SWD）切片或塑料培养皿上并连续培养，或每周或每半周用胰蛋白酶消化并重新接种到新鲜基质上。通过光学显微镜和扫描电子显微镜进行观察。沉降后立即在骨髓部分观察到许多多核细胞，推测它们源自紧邻骨的附着性差的破骨细胞，这些破骨细胞在收获时未进行吸收。在塑料上培养三天后，多核细胞在平面范围和核数量方面都非常大：破骨细胞之间以及破骨细胞与小的圆形单核细胞之间发生了细胞融合。SWD被大量吸收。（a）胰蛋白酶消化后，破骨细胞对塑料的黏附性比其他细胞更强；（b）对SWD的黏附性比对塑料更强，尤其是当细胞正在进行吸收时。胰蛋白酶消化后的细胞接种到新的SWD上后恢复了吸收能力，但仅能进行1或2次处理。除了其他骨细胞类型的比例高得多之外，骨来源的细胞与骨髓培养物相似。胰蛋白酶消化导致SWD和塑料上的多核细胞比例损失更高。所有培养物在6周时仍在进行吸收。吸收坑的形状、深度、平面面积和体积存在广泛差异。
