Brighton C T, Shadle C A, Jimenez S A, Irwin J T, Lane J M, Lipton M
Arthritis Rheum. 1979 Oct;22(10):1093-101. doi: 10.1002/art.1780221008.
Articular cartilage slice explants were stored under various conditions, including freezing-thawing at various rates by using dimethyl sulfoxide (DMSO) as a cryoprotective agent, incubating in standard tissue culture medium (MEM Eagle:NCTC 135:15% fetal calf serum) in 5% CO2 and air at 4 degrees, 21 degrees, and 37 degrees C, and incubating in standard tissue culture medium containing 200 micrograms/ml alpha-tocopherol (vitamin E) at 37 degrees C after first ascertaining a dose-response curve of vitamin E. Results indicated that articular cartilage slice explants did not survive freezing or storage at 4 degrees and 21 degrees C as measured by 35S uptake. When stored at 37 degrees C in standard tissue culture in 5% CO2 and air, the slice explants remained viable for up to 60 days. The addition of alpha-tocopherol to the medium resulted in significantly less release of previously incorporated 35Sin stored cartilage slices and significantly less reduction of the amount of hexosamine present in the stored explants. alpha-Tocopherol in the medium also preserved safranin O staining. Thus, the application of tissue culture techniques to the storage of articular cartilage made it possible to preserve cartilage slice explants in a viable, biochemically "normal" state.
关节软骨切片外植体在多种条件下保存,包括使用二甲基亚砜(DMSO)作为冷冻保护剂以不同速率进行冻融,在含5%二氧化碳和空气的标准组织培养基(伊格尔氏MEM培养基:NCTC 135培养基:15%胎牛血清)中于4℃、21℃和37℃孵育,以及在首先确定维生素E的剂量反应曲线后,于37℃在含200微克/毫升α-生育酚(维生素E)的标准组织培养基中孵育。结果表明,通过35S摄取量测定,关节软骨切片外植体在4℃和21℃冷冻或保存时无法存活。当在含5%二氧化碳和空气的标准组织培养基中于37℃保存时,切片外植体可存活长达60天。向培养基中添加α-生育酚可显著减少储存软骨切片中先前掺入的35S的释放,并显著减少储存外植体中己糖胺含量的降低。培养基中的α-生育酚还能保持番红O染色。因此,将组织培养技术应用于关节软骨的储存,使得软骨切片外植体能够在存活、生化“正常”的状态下得以保存。