A novel virally inactivated human platelet lysate preparation rich in TGF-β, EGF and IGF, and depleted of PDGF and VEGF
A novel virally inactivated human platelet lysate preparation rich in TGF-β, EGF and IGF, and depleted of PDGF and VEGF
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DOI：
10.1042/ba20100151
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发表时间：
2010-08-01
影响因子：
2.8
通讯作者：
Burnouf, Thierry
中科院分区：
文献类型：
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作者：
Burnouf, Pierre-Alain;Juan, Po-Kai;Burnouf, Thierry
在细胞培养、细胞治疗和临床应用中，人们对使用富含GFS(生长因子)的标准化的病毒灭活的人血小板裂解制剂产生了兴趣。本文报道了一种简单的制备病毒灭活的血小板裂解物制剂的方法，该制剂富含转化生长因子-β1(TGF-β1)、表皮生长因子(EGF)和胰岛素样生长因子(IGF)，而缺乏血小板衍生生长因子(PDGF)和血管内皮生长因子(VEGF)。用S/D(溶剂/洗涤剂)病毒灭活法处理浓缩血小板，然后用油提取，然后用活性碳吸附，最后无菌过滤。所得制剂中转化生长因子-β1、表皮生长因子和胰岛素样生长因子的平均含量分别为368.4、2.4和54.7 ng/ml。经过炭化处理后，PDGF-AB和血管内皮生长因子基本上被完全去除。白蛋白、Ig G、Ig M、Ig A和纤维蛋白原含量的平均值接近。分别为40.0、8.5、0.87、1.66和2.65 mg/ml，胆固醇和甘油三酯分别为15和20.7 mg/ml，磷酸三正丁酯和Triton X-45分别为8.7和8.8。分别进行了分析。在最低必需培养液中添加1-10%的S/D处理的血小板裂解液，可促进MG63和SiRC细胞的增殖，并促进或优于10%(v/v)胎牛血清的增殖[3-(4，dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)2-(4-sulfophenyl)-2H-tetrazolium]法)]。用于制备这种经S/D处理的血小板裂解物的工艺易于工业生产。我们的结果为评估这种新制剂在干细胞扩增和/或骨组织工程和再生方面的潜力提供了可能。
There is emerging interest in the use of standardized virally inactivated human platelet lysate preparations rich in GFs (growth factors) for cell cultures, cell therapy and clinical applications. In the present paper, we report a simple process to prepare a virally inactivated platelet lysate preparation rich in TGF-beta 1 (transforming growth factor-beta 1), EGF (epidermal growth factor) and IGF (insulin-like growth factor) and depleted of PDGF (platelet-derived growth factor) and VEGF (vascular endothelial growth factor). Apheresis platelet concentrates were treated by the S/D (solvent/detergent) viral inactivation procedure, then subjected to an oil extraction followed by adsorption with activated charcoal and finally sterile-filtered. The resulting preparation contained a mean of 368.4, 2.4 and 54.7 ng/ml of TGF-beta 1, EGF and IGF respectively. PDGF-AB and VEGF were essentially completely removed by the charcoal treatment. The mean albumin, IgG, IgM and IgA and fibrinogen contents were approx. 40.0, 8.5, 0.87, 1.66 and 2.65 mg/ml respectively, cholesterol and triglycerides were at 15 and 20.7 mg/ml respectively and TnBP (tri-n-butyl phosphate) and Triton X-45 were at 8.7 and 8.8 p.p.m. respectively. Supplementing MEM (minimum essential medium) with 1-10% of this S/D-treated platelet lysate promoted the proliferation of MG63 and SIRC cell lines as well as, or better than, 10% (v/v) FBS (fetal bovine serum), as based on the MTS [3-(4, dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)2-(4-sulfophenyl)-2H-tetrazolium] assay. The process used to prepare such S/D-treated platelet lysates is easily scalable for industrial production. Our results open up the possibility to evaluate the potential of this new preparation for stem cell expansion and/or bone tissue engineering and regeneration.