Schallmoser Katharina, Bartmann Christina, Rohde Eva, Reinisch Andreas, Kashofer Karl, Stadelmeyer Elke, Drexler Camilla, Lanzer Gerhard, Linkesch Werner, Strunk Dirk
Department of Blood Group Serology and Transfusion Medicine, Division of Hematology and Stem Cell Transplantation, Medical University, Auenbrugger Platz 38, A-8036 Graz, Austria.
Transfusion. 2007 Aug;47(8):1436-46. doi: 10.1111/j.1537-2995.2007.01220.x.
Human multipotent mesenchymal stromal cells (MSCs) are promising candidates for a growing spectrum of regenerative and immunomodulatory cellular therapies. Translation of auspicious experimental results into clinical applications has been limited by the dependence of MSC propagation from fetal bovine serum (FBS).
The capacity of human platelet lysate (HPL) to replace FBS for clinical-scale MSC propagation was analyzed.
HPL could be efficiently produced from buffy coats. Multiplex analyses allowed a distinct HPL growth factor profile to be delineated. With a previously established two-step clinical-scale procedure, HPL was reproducibly more efficient than FBS in supporting MSC outgrowth. With only 3 x 10(5) primary culture-derived MSCs, a mean of 4.36 x 10(8) HPL-MSCs (range, 3.01 x 10(8)-5.40 x 10(8)) was obtained within a single secondary 11- to 13-day culture step. Although morphologically distinct, HPL-MSCs and FBS-MSCs did not differ significantly in terms of immunophenotype, differentiation potential in vitro, and lack of tumorigenicity in nude mice in vivo.
Replacing FBS with HPL prevents bovine prion, viral, and zoonose contamination of the stem cell product. This new efficient FBS-free two-step procedure for clinical-scale MSC propagation may represent a major step toward challenging new stem cell therapies.
人多能间充质基质细胞（MSC）是越来越多再生和免疫调节细胞疗法中很有前景的候选细胞。由于MSC的增殖依赖胎牛血清（FBS），使得将良好的实验结果转化为临床应用受到限制。
分析了人血小板裂解液（HPL）替代FBS用于临床规模MSC增殖的能力。
HPL可从血沉棕黄层高效制备。多重分析能够描绘出独特的HPL生长因子谱。采用先前建立的两步临床规模程序，在支持MSC生长方面，HPL比FBS更高效且可重复。仅用3×10⁵原代培养的MSC，在单个11至13天的二级培养步骤中，平均可获得4.36×10⁸个HPL-MSC（范围为3.01×10⁸至5.40×10⁸）。尽管形态上有差异，但HPL-MSC和FBS-MSC在免疫表型、体外分化潜能以及体内裸鼠致瘤性方面无显著差异。
用HPL替代FBS可防止干细胞产品受到牛朊病毒、病毒和人畜共患病原的污染。这种用于临床规模MSC增殖的新型高效无FBS两步程序可能是迈向新型干细胞疗法的重要一步。
