目的 了解广州地区已育女性献血者中HLA-Ⅰ和Ⅱ类抗体的分布和特异性,评估这类献血者血液及其成分所含HLA抗体引起免疫性输血不良反应的风险.方法 随机收集广州地区已育女性献血者标本343(人)份、未育女性献血者标本140(人)份,利用Lifecodes LifeScreen Deluxe(LMX)筛查标本的HLA抗体,阳性者再分别利用LIFECODES LSATMClass Ⅰ和LIFECODES LSATMClassⅡ鉴定抗体特异性.结果 广州地区已育女性献血者中,抗-HLA阳性率25.07％ (86/343),而未育女性献血者中抗-HLA阳性率3.57％ (5/140) (P ＜0.05).已育女性献血者中HLA-Ⅰ类抗体出现频率较高的分别是抗-A* 11 28.8％ (17/59)、抗-B* 07 42.4％ (25/59)和抗-Cw* 07 20.3％(12/59),HLA-Ⅱ类抗体出现频率最高者是抗-DRB1 *04 40.4％ (19/47).结论 已育女性献血者HLA抗体出现的频率较高,这类献血者的血浆和单采血小板用于临床后势必增加HLA同种免疫反应的风险,应引起高度重视.

Objective To provide an efficient, practicable and high specificity method for culture of bovine retinal capillary endothelial cells(BRCEC). Methods Fresh bovine eyes were obtained from a local market. The retinas were removed and washed several times in DMEM. Subsequently retinas were homogenized and the resultant pellet was resuspended in an isolation medium. Microvesseles were trapped on an 75 μm nylon mesh and transferred to a 50 ml centrifuge tube containing 10 ml enzyme cocktail which includes collagenase Ⅰ, DNase Ⅰand pronase and treated for 20 min. The resultant vessel fragments were trapped on a 46 μm nylon mesh,washed with the isolation medium and centrifuged for 5 min. For selective culture of BRCECs, the resultant pellet was resuspended in 2 ml of the BRCEC growth medium and transferred into 25 cm2 gelatin-coated plastic tissue culture flasks. BRCECs were identified by immunohistochemical method of factor Ⅷ. Results The purity of selectively cultured BRCECs was more than 95%. Cells seeded onto the surface of the polymerized EC Matrix could develop the cellular network structures by 12-18 h. Conclusion BRCECs can be attained easily and effectively by this method. This culture method would provide a useful model for studying disease related to retinal vessels in vitro.