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GMP‐grade CD34 + selection from HLA‐homozygous licensed cord blood units and short‐term expansion under European ATMP regulations
Author(s) -
Liedtke Stefanie,
Korschgen Lutz,
Korn Janine,
Duppers Almuth,
Kogler Gesine
Publication year - 2021
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/vox.12978
Subject(s) - cord blood , cd34 , population , medicine , andrology , immunology , chemistry , stem cell , biology , genetics , environmental health
Background Based on a synergistic consortium, the cord blood (CB) bank Düsseldorf 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 Düsseldorf 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 10 5 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 Düsseldorf 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.