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Manufacture of endothelial colony‐forming progenitor cells from steady‐state peripheral blood leukapheresis using pooled human platelet lysate
Author(s) -
Siegel Georg,
Fleck Erika,
Elser Stefanie,
HermanutzKlein Ursula,
Waidmann Marc,
Northoff Hinnak,
Seifried Erhard,
Schäfer Richard
Publication year - 2018
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.14541
Subject(s) - leukapheresis , immunology , andrology , population , cd34 , vascular endothelial growth factor , cd31 , progenitor cell , apheresis , matrigel , platelet , biology , medicine , angiogenesis , microbiology and biotechnology , stem cell , cancer research , environmental health , vegf receptors
BACKGROUND Endothelial colony‐forming progenitor cells (ECFCs) are promising candidates for cell therapies. However, ECFC translation to the clinic requires optimized isolation and manufacture technologies according to good manufacturing practice (GMP). STUDY DESIGN AND METHODS ECFCs were manufactured from steady‐state peripheral blood (PB) leukapheresis (11 donors), using GMP‐compliant technologies including pooled human platelet (PLT) lysate, and compared to human umbilical cord endothelial cells, human aortic endothelial cells, and human cerebral microvascular endothelial cells. Specific variables assessed were growth kinetics, phenotype, trophic factors production, stimulation of tube formation, and Dil‐AcLDL uptake. RESULTS ECFCs could be isolated from PB leukapheresis units with mean processed volume of 5411 mL and mean white blood cell (WBC) concentration factor of 8.74. The mean frequency was 1.44 × 10 −8 ECFCs per WBC, corresponding to a mean of 177.8 ECFCs per apheresis unit. Expandable for up to 12 cumulative population doublings, calculated projection showed that approximately 730 × 10 3 ECFCs could be manufactured from 1 apheresis unit. ECFCs produced epidermal growth factor, hepatocyte growth factor, vascular endothelial growth factor (VEGF)‐A, PLT‐derived growth factor‐B, interleukin‐8, and monocyte chemoattractant protein‐1, featured high potential for capillary‐like tubes formation, and showed no telomerase activity. They were characterized by CD29, CD31, CD44, CD105, CD117, CD133, CD144, CD146, and VEGF‐R2 expression, with the most common subpopulation CD34+CD117–CD133–. Compared to controls, ECFCs featured greater Dil‐AcLDL uptake and higher expression of CD29, CD31, CD34, CD44, CD144, and VEGF‐R2. CONCLUSIONS Here we show that isolation of ECFCs with proangiogenic profile from steady‐state PB leukapheresis is feasible, marking a first step toward ECFC product manufacture according to GMP.