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| Item Type: | Article |
|---|---|
| Title: | Widely applicable, extended flow cytometric stem cell enumeration panel for quality control of advanced cellular products |
| Creators Name: | Haussmann, K. and Streitz, M. and Takvorian, A. and Grund, J. and Skenderi, Z. and Tietze-Bürger, C. and Movassaghi, K. and Künkele, A. and Blum, A. and Bullinger, L. |
| Abstract: | The most widely used quality control assay for CD34 + hematopoietic stem cell product characterization is the protocol established by the International Society of Hematotherapy and Graft Engineering (ISHAGE). While this protocol is still the gold standard for stem cell enumeration and viability assessment, it does not include T cell enumeration, which is nowadays mandatory for assaying standard allogeneic grafts and various advanced therapy medicinal products (ATMPs). In accordance, we have developed and extensively validated a new approach for a more comprehensive characterization of hematopoietic cellular products using a pre-formulated dried antibody format panel. In addition to the counting beads, the typical markers CD45 fluorescein isothiocyanate (FITC) and CD34 phycoerythrin (PE), as well as the viability dye 7-amino actinomycin D (7-AAD), our novel pre-formulated panel also contains CD3 Pacific Blue (PB) and CD19 allophycocyanin (APC) in the same tube, thereby allowing a combined calculation of leucocytes, stem cells, T and B cells. Showing high linearity, sensitivity and accuracy, our approach is easy to implement and enables a more in-depth characterization of the cellular product under release testing conditions. In addition, the dried pre-formulated antibody approach increases assay reliability compared to the standard antibody panel. |
| Keywords: | CD34 Antigens, Flow Cytometry, Fluorescein-5-isothiocyanate, Hematopoietic Stem Cells, Phycoerythrin, Quality Control, Reproducibility of Results |
| Source: | Scientific Reports |
| ISSN: | 2045-2322 |
| Publisher: | Nature Publishing Group |
| Volume: | 12 |
| Number: | 1 |
| Page Range: | 17995 |
| Date: | 26 October 2022 |
| Official Publication: | https://doi.org/10.1038/s41598-022-22339-1 |
| PubMed: | View item in PubMed |
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