GMP‐grade CD34 + selection from HLA‐homozygous licensed cord blood units and short‐term expansion under European ATMP regulations
| Autor: | Almuth Duppers, Janine Korn, Stefanie Liedtke, Lutz Korschgen, Gesine Kögler |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
education.field_of_study
Manufacturing process business.industry Population Hematology General Medicine Human leukocyte antigen 030204 cardiovascular system & hematology Andrology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine chemistry Cord blood Cd34 selection Medicine ATMP education business 030215 immunology |
| Zdroj: | Vox Sanguinis. 116:123-135 |
| ISSN: | 1423-0410 0042-9007 |
| DOI: | 10.1111/vox.12978 |
| Popis: | Background Based on a synergistic consortium, the cord blood (CB) bank Dusseldorf was responsible for the selection of HLA-homozygous (HLA-h) donors, contacting/re-consenting the mothers, Good Manufacturing Practice (GMP)-grade CD34+ enrichment, followed by short-term expansion of CD34+ cells and qualification of the resulting CD34+ population as advanced therapy medicinal product (ATMP)-starting material. Among 20 639 licensed Dusseldorf cord blood units (CBUs), 139 potential HLA-h donors were identified with the most frequent 10 German haplotypes. 100% of the donors were contacted, and for 47·5%, consent was obtained. HLA-A, -B, -C, -DR, -DQ and -DP were determined by sequencing. Methods Thawing/washing of the CBUs was performed in the presence of Volulyte/HSA with Sepax® , CD34+ selection by automated CliniMACS® -system (Miltenyi), expansion with qualified GMP-grade cytokines and media in the GMP facility. Results Here, we specify minimal criteria (≥5 x 105 viable CD34+ -count, ≥80% CD34+ -purity and ≥70% viability) and confirm that n = 10 CB units (max storage time 16 years) could be qualified for an ATMP starting material. The mean fold change expansion of isolated CD34+ cells at Day 3/4 (d3/4) was 3·38 ± 3·02 with a mean purity of 86·90 ± 10·38% and a high viability of 96·07 ± 4·72%. Conclusion As of March 2019, approval was obtained by the Bezirksregierung Dusseldorf for the GMP-compliant production. The production of HLA-homozygous expanded CD34+ cells from cryopreserved CB under European ATMP regulations presented here describes the successful clinical translation and implementation of a qualified manufacturing process. This approach considers the main obstacle of rejection of transplanted cells (due to the immunological HLA barrier) by preselection of HLA-homozygous transplants. |
| Databáze: | OpenAIRE |
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