The monoculture of cord-blood-derived CD34+ cells by an automated, membrane-based dynamic perfusion system with a novel cytokine cocktail

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 × 106 positively selected CD34+ cell inoculum from 3 donor lineages, the mean cell harvest and cell viability were 1.02 × 108 cells and 95.5%, respectively, and the mean frequency of the CD45+CD34+ 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.

 

Comments：

It appears that you have provided information about an automated expansion protocol for cord blood-derived CD34+ cells. This protocol uses a novel cytokine cocktail and a fibronectin-stromal-cell-derived factor-1 (SDF-1)-coated membrane in a dynamic perfusion bioreactor called the Quantum system. The aim of this protocol is to provide a sufficient number of T cell-depleted hematopoietic stem and progenitor cells for use in human leukocyte antigen (HLA)-matched cord blood (CB) transplantation to treat certain hematological malignancies, hemoglobinopathies, and autoimmune disorders.

According to the results of an 8-day expansion of a 2 × 106 positively selected CD34+ cell inoculum from 3 donor lineages, the mean cell harvest and cell viability were 1.02 × 108 cells and 95.5%, respectively. The mean frequency of the CD45+CD34+ 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 suggest that the automated monoculture protocol can support the expansion of CD34+ cells with minimal lymphocyte residual.

Overall, this information suggests that this automated expansion protocol could be a useful tool in providing a sufficient number of T cell-depleted hematopoietic stem and progenitor cells for use in HLA-matched CB transplantation.

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