An Injectable Solution for Preservation of Hematopoietic Stem and Progenitors Cells in Hypothermic Condition.

Abstract

To ensure the preservation of functional hematopoietic stem cells (HSC) and committed progenitor cells (HPC) at + 4 °C in ex vivo expanded cord blood cell products during worldwide transportation and subsequent infusion-without the need for washing and cell concentration-we developed a conservation medium called Stabilizer of Expanded Cells (SEC), composed exclusively of injectable pharmacological products. The in vivo engraftment assay in immunodeficient mice was used to detect primitive HSCs before and after preservation at + 4 °C. In some experiments, a complex phenotype based on CD34, CD38, and CD133 expression was utilized for this purpose. Committed progenitors (CFU-GM, BFU-E, and CFU-Mix) were detected using methylcellulose culture colony-forming assays. Additionally, in some cases, the energetic metabolism (mitochondrial respiration) was evaluated using Seahorse technology. SEC was able to preserve the functionality of HSCs and HPCs in ex vivo expanded cell populations at + 4 °C for at least 48 h. Furthermore, SEC is also effective in fully preserving HSCs and HPCs in cytapheresis products for at least 72 h. Additionally, SEC enabled the full preservation of HSCs and HPCs for 72 h in freshly collected cord blood, maintaining a normal metabolic profile of CD34⁺ cells. The SEC medium exhibits a positive effect on the maintenance of both HSCs and HPCs at + 4 °C, regardless of their source. Therefore, SEC can be applied in cell therapy protocols based on HSCs and HPCs with a significant advantage: the product does not need to be washed and concentrated before injection into the patient.