Research and Development, Terumo Blood and Cell Technologies, Lakewood, CO 80215, USA.
Research and Development, Terumo Blood and Cell Technologies, Lakewood, CO 80215, USA.
Stem Cell Reports. 2022 Dec 13;17(12):2585-2594. doi: 10.1016/j.stemcr.2022.10.006. Epub 2022 Nov 3.
Human leukocyte antigen (HLA)-matched cord blood (CB) transplantation is a procedure for the treatment of certain hematological malignancies, hemoglobinopathies, and autoimmune disorders. However, one of the challenges is to provide a sufficient number of T cell-depleted hematopoietic stem and progenitor cells. Currently, only 4%-5% of the CB units stored in CB banks contain enough CD34 cells for engrafting 70-kg patients. To support this clinical need, we have developed an automated expansion protocol for CB-derived CD34 cells in the Quantum system's dynamic perfusion bioreactor using a novel cytokine cocktail comprised of stem cell factor (SCF), thrombopoietin (TPO), fms-like tyrosine kinase 3 ligand (Flt-3L), interleukin-3 (IL-3), IL-6, glial cell line-derived neurotrophic factor (GDNF), StemRegenin 1 (SR-1), and a fibronectin-stromal-cell-derived factor-1 (SDF-1)-coated membrane. In an 8-day expansion of a 2 × 10 positively selected CD34 cell inoculum from 3 donor lineages, the mean cell harvest and cell viability were 1.02 × 10 cells and 95.5%, respectively, and the mean frequency of the CD45CD34 immunophenotype was 54.3%. The mean differentiated cell frequencies were 0.5% for lymphocytes, 15.8% for neutrophils, and 15.4% for platelets. These results demonstrate that the automated monoculture protocol can support the expansion of CD34 cells with minimal lymphocyte residual.
人类白细胞抗原 (HLA) 匹配的脐带血 (CB) 移植是治疗某些血液恶性肿瘤、血红蛋白病和自身免疫性疾病的一种方法。然而，其中一个挑战是提供足够数量的 T 细胞 depleted 造血干细胞和祖细胞。目前，CB 银行储存的 CB 单位中只有 4%-5% 含有足够的 CD34 细胞，可植入 70-kg 患者。为了满足这一临床需求，我们在 Quantum 系统的动态灌注生物反应器中使用一种由干细胞因子 (SCF)、血小板生成素 (TPO)、fms 样酪氨酸激酶 3 配体 (Flt-3L)、白细胞介素-3 (IL-3)、白细胞介素-6 (IL-6)、胶质细胞衍生神经营养因子 (GDNF)、StemRegenin 1 (SR-1) 和纤维连接蛋白基质细胞衍生因子-1 (SDF-1) 涂层膜组成的新型细胞因子鸡尾酒，开发了一种用于 CB 衍生 CD34 细胞的自动化扩增方案。在对来自 3 个供体谱系的 2×10 个正选择 CD34 细胞接种物进行 8 天的扩增中，平均细胞收获量和细胞活力分别为 1.02×10 个细胞和 95.5%，CD45CD34 免疫表型的平均频率为 54.3%。分化细胞的平均频率分别为淋巴细胞 0.5%、中性粒细胞 15.8%和血小板 15.4%。这些结果表明，自动化的单核细胞培养方案可以在最小淋巴细胞残留的情况下支持 CD34 细胞的扩增。
