Two-dimensional polymer-based cultures expand cord blood-derived hematopoietic stem cells and support engraftment of NSG mice.
Currently, ex vivo expansion of hematopoietic stem cells (HSC) is still insufficient. Traditional approaches for HSC expansion include the use of stromal cultures, growth factors, and/or bioreactors. Biomaterial-based strategies provide new perspectives. We focus on identifying promising two-dimensional (2D) polymer candidates for HSC expansion. After a 7-day culture period with cytokine supplementation, 2D fibrin, poly(D,L-lactic-co-glycolic acid; Resomer® RG503), and Poly(ɛ-caprolactone; PCL) substrates supported expansion of cord blood (CB)-derived CD34⁺ cells ex vivo. Fibrin cultures achieved the highest proliferation rates (>8700-fold increase of total nuclear cells, p<0.001), high total colony-forming units (3.6-fold increase, p<0.001), and highest engraftment in NSG mice (7.69-fold more donor cells compared with tissue culture polysterene, p<0.001). In addition, the presence of multiple human hematopoietic lineages such as myeloid (CD13⁺), erythroid (GypC⁺), and lymphoid (CD20⁺/CD56⁺) in murine transplant recipients confirmed the multilineage engraftment potential of fibrin-based cultures. Filopodia development in fibrin-expanded cells was a further indicator for superior cell adhesion capacities. We propose application of fibrin, Resomer® RG503, and PCL for future strategies of CB-CD34⁺ cell expansion. Suitable polymers for HSC expansion might also be appropriate for future drug discovery applications or for studies aimed to develop hematological therapies.
Institute of Pathology, RWTH Aachen University, Aachen, Germany. mventuraferreira@ukaachen.de
Mónica Sofia Ventura
MS
Ferreira
Rebekka Kramann
RK
Schneider
Wolfgang
W
Wagner
Willi
W
Jahnen-Dechent
Norina
N
Labude
Manfred
M
Bovi
Daniela
D
Piroth
Ruth
R
Knüchel
Thomas
T
Hieronymus
Albrecht M
AM
Müller
Martin
M
Zenke
Sabine
S
Neuss
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape; D002470:Cell Survival; D005312:Fetal Blood / Q000166:cytology*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D053631:Interleukin Receptor Common gamma Subunit / Q000172:deficiency / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009928:Organ Specificity / Q000187:drug effects; D011108:Polymers / Q000494:pharmacology*; D014410:Tumor Stem Cell Assay
D018952:Antigens, CD34; D015415:Biomarkers; C507942:Il2rg protein, mouse; D053631:Interleukin Receptor Common gamma Subunit; D011108:Polymers