Human hematopoietic stem cells (HSCs) are widely used as a cellular source for hematopoietic stem cell transplantation (HSCT) in the clinical treatment of hematological malignancies. After transplantation therapy, delays in hematopoietic recovery due to insufficient donor-derived HSCs can lead to increased risks of life-threatening infections and bleeding. Our previous studies developed an efficient ex vivo expansion culture medium (3a medium) for umbilical cord blood-derived HSCs (CBSCs), offering a potential solution to this problem. Nevertheless, the broader applicability of our culture method to alternative cell sources and, of greater significance, its efficacy in eliminating potentially disease-associated contaminated tumor cells, especially in autologous transplantation, raise critical clinical questions. In this study, we modified the 3a medium by incorporating UM729 to replace UM171, adding FMS-like tyrosine kinase 3 (Flt3) ligand, and adjusting the concentrations of butyzamide, 740Y-P, polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (PCL-PVAc-PEG, Soluplus) to create the modified-3a medium. This sophistication allowed the efficient expansion of not only CBSCs but also peripheral blood-mobilized HSCs (PBSCs). Additionally, we successfully removed contaminated myeloma cells by adding bortezomib and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) at appropriate concentrations, although we maintained HSCs through the addition of lenalidomide. Our research findings present the potential for widespread clinical application of the modified-3a medium and suggest a safe ex vivo culture technique for expanding human HSCs within peripheral blood-derived donor grafts used for autologous HSCT.

中文翻译：

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清除骨髓瘤细胞污染物并同时扩增外周血动员干细胞

人造血干细胞（HSC）作为造血干细胞移植（HSCT）的细胞来源广泛应用于血液系统恶性肿瘤的临床治疗。移植治疗后，由于供体来源的造血干细胞不足而导致造血恢复延迟，可能导致危及生命的感染和出血的风险增加。我们之前的研究开发了一种高效的脐带血源性造血干细胞（CBSC）离体扩增培养基（3a培养基），为这一问题提供了潜在的解决方案。然而，我们的培养方法对替代细胞来源的更广泛适用性，更重要的是，它在消除潜在与疾病相关的污染肿瘤细胞方面的功效，特别是在自体移植中，提出了关键的临床问题。在本研究中，我们修改了3a培养基，加入UM729代替UM171，添加FMS样酪氨酸激酶3（Flt3）配体，并调整丁酰胺、740Y-P、聚乙烯己内酰胺-聚乙酸乙烯酯-聚乙二醇接枝共聚物的浓度（ PCL-PVAc-PEG、Soluplus）来创建改良的 3a 培养基。这种复杂性不仅可以有效扩增 CBSC，还可以有效扩增外周血动员的 HSC (PBSC)。此外，我们通过添加适当浓度的硼替佐米和肿瘤坏死因子（TNF）相关凋亡诱导配体（TRAIL）成功去除了污染的骨髓瘤细胞，尽管我们通过添加来那度胺维持了HSC。我们的研究结果展示了改良 3a 培养基广泛临床应用的潜力，并提出了一种安全的离体培养技术，用于在用于自体 HSCT 的外周血来源供体移植物中扩增人类 HSC。