Goodrich Raymond P, Li Junzhi, Pieters Henry, Crookes Robert, Roodt Jan, Heyns Anthon du P
Navigant Biotechnologies, Inc., Lakewood, CO 80215, USA.
Vox Sang. 2006 May;90(4):279-85. doi: 10.1111/j.1423-0410.2006.00761.x.
Changes in in vitro platelet quality parameters during platelet storage are associated with a decrease of in vivo platelet viability after platelet transfusion. Many attempts have been made to identify the most predictable in vitro parameters for in vivo performance. We used a riboflavin-based ultraviolet (UV) light treatment process designed to inactivate pathogens and white blood cell (WBC) contaminants in blood products as a model system in which to study the correlation of in vitro cell quality with in vivo viability.
Platelet products (n = 18) were collected by a standard Trima apheresis procedure and treated with one of three dose levels of UV light (0, 7.2 or 12.4 J/ml) in the presence of 50 microm riboflavin. Lactate production, glucose consumption and P-selectin expression, pH, pCO(2), pO(2), hypotonic shock response and swirl were measured during 5 days of platelet storage post-UV/RB treatment. Aliquots of these products were radiolabelled on day 5 of storage and were subsequently used to determine platelet recovery and survival time in autologous subjects.
The responses of in vitro cell quality were observed to occur in a UV dose-dependent manner. Lactate production and pH were identified as the parameters most strongly correlated with platelet in vivo recovery, which ranged from 5 to 82%. The correlation coefficients (r) for lactate production and pH with in vivo recovery in human subjects were 0.9090 and 0.8831 with P-values of 0.007 and 0.031, respectively. Lactate production and pH were also found to be correlated with platelet survival time, with correlation coefficients of 0.8063 and 0.8384 (the P values were 0.01 and 0.001, respectively).
Using conditions of riboflavin-based UV light treatment, lactate production and pH were identified as having the highest correlations with recovery and survival of radiolabelled platelets in healthy subjects.
血小板储存期间体外质量参数的变化与血小板输注后体内活力的降低有关。人们进行了许多尝试,以确定对体内性能最具预测性的体外参数。我们使用一种基于核黄素的紫外线(UV)光处理工艺,该工艺旨在灭活血液制品中的病原体和白细胞(WBC)污染物,作为一个模型系统来研究体外细胞质量与体内活力之间的相关性。
通过标准的Trima血液成分单采程序收集血小板制品(n = 18),并在存在50微摩尔核黄素的情况下,用三种剂量水平的紫外线(0、7.2或12.4 J/ml)之一进行处理。在紫外线/核黄素处理后的血小板储存5天期间,测量乳酸生成、葡萄糖消耗、P-选择素表达、pH值、pCO₂、pO₂、低渗休克反应和涡旋情况。这些制品的等分试样在储存第5天进行放射性标记,随后用于确定自体受试者体内的血小板回收率和存活时间。
观察到体外细胞质量的反应呈紫外线剂量依赖性。乳酸生成和pH值被确定为与体内血小板回收率相关性最强的参数,回收率范围为5%至82%。在人类受试者中,乳酸生成和pH值与体内回收率的相关系数(r)分别为0.9090和0.8831,P值分别为0.007和0.031。还发现乳酸生成和pH值与血小板存活时间相关,相关系数分别为0.8063和0.8384(P值分别为0.01和0.001)。
在基于核黄素的紫外线光处理条件下,乳酸生成和pH值被确定为与健康受试者体内放射性标记血小板的回收率和存活率相关性最高。