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Comparison of human platelet lysate alternatives using expired and freshly isolated platelet concentrates for adipose-derived stromal cell expansion

机译:使用过期和新鲜分离的血小板浓缩物进行人血小板裂解物替代品的脂肪衍生的基质细胞膨胀的比较

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Pooled human platelet lysate (pHPL) has been used to expand adipose-derived stromal cells (ASCs) and can be formulated using fresh or expired buffy coats (BCs) which are then resuspended in either plasma or an additive solution. Not much is known about the effects that expired products and additive solutions have on ASC expansion, and the need for quality control and release criteria has been expressed. This pilot study compared proliferation, cell size, morphology and immunophenotype of ASCs expanded in the different pHPL alternatives versus foetal bovine serum (FBS). Quality control criteria were assessed prior to and during the manufacture of the pHPL alternatives. ASCs were then expanded in 1%, 2.5%, 5% or 10% of the different pHPL alternatives or in 10% FBS. Cell size, morphology, cell number and immunophenotype were measured using microscopy and flow cytometry. The majority of the pHPL alternatives were within the recommended ranges for the quality control criteria. ASCs expanded in the pHPL alternatives were smaller in size, displayed a tighter spindle-shaped morphology, increased cell growth and had a similar immunophenotype (with the exception of CD34 and CD36) when compared to ASCs expanded in FBS. Here we report on the effects that expired BC products and additive solutions have on ASC expansion. When taken together, our findings indicate that all of the pHPL alternatives can be considered to be suitable replacements for FBS for ASC expansion, and that expired BC products can be used as an alternative to fresh BC products.
机译:合并的人血小板裂解物(PHPL)已被用于扩增脂肪衍生的基质细胞(ASCS),并且可以使用新鲜或过期的乳腺涂层(BCS)配制,然后在血浆或添加剂溶液中重悬。对于已知的产品和添加剂溶液对ASC扩展的影响并不多,并且已经表达了对质量控制和释放标准的需求。该试点研究比较了不同PHPL替代品与胎牛血清(FBS)扩增的ASC的增殖,细胞大小,形态和免疫蛋白酶型。在制造PHPL替代品之前和期间评估质量控制标准。然后在不同PHPL替代品的1%,2.5%,5%或10%的1%,2.5%,5%或10%中扩增ASC。使用显微镜和流式细胞术测量细胞尺寸,形态,细胞数和免疫蛋白型。大多数PHPL替代品在建议的质量控制标准范围内。在PHPL替代方案中扩张的尺寸较小,呈现更严格的主轴形状态,增加细胞生长,并且与在FBS中膨胀时,具有类似的免疫表型(除CD34和CD36外)。在这里,我们报告了已过期的BC产品和添加剂解决方案对ASC扩展的影响。当占用时,我们的研究结果表明,所有PHPL替代方案都可以被认为是ASC扩展的FBS合适的替代品,并且过期的BC产品可用作新鲜BC产品的替代品。

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