TY - JOUR

T1 - A parallelized, perfused 3D triculture model of leukemia for in vitro drug testing of chemotherapeutics

AU - Zippel, Sabrina

AU - Dilger, Nadine

AU - Chatterjee, Chandralekha

AU - Raic, Annamarija

AU - Brenner-Weiß, Gerald

AU - Schadzek, Patrik

AU - Rapp, Bastian E.

AU - Lee-Thedieck, Cornelia

N1 - Funding Information: This work was supported by the NanoMatFutur program of the German Federal Ministry of Education and Research (BMBF; FKZ 13N12968 and 13XP5076A). This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (Grant Agreement No. 757490). This work also received support from the framework of the SMART BIOTECS alliance between the Technische Universität Braunschweig and the Leibniz Universität Hannover. This initiative is supported by the Ministry of Science and Culture (MWK) of Lower Saxony, Germany. The authors thank Saskia Kraus (Institute of Functional Interfaces, KIT and Institute of Cell Biology and Biophysics, Leibniz University Hannover) for her excellent technical assistance and support in the preparation of magnetic hydrogels, Dr. Domenic Kratzer (Institute of Functional Interfaces, KIT and Institute of Cell Biology and Biophysics, Leibniz University Hannover) for SEM imaging, Frank Kirschhöfer (Institute of Functional Interfaces, KIT) and Michael Nusser (Institute of Functional Interfaces, KIT) for the support regarding adenosine and amino acid determination, PD Dr. Frank Schaarschmidt (Institute of Cell Biology and Biophysics, Leibniz University Hannover) for statistical consultation and Kai Sachsenheimer (Institute of Microstructure Technology, KIT) for rebuilding of the magnet lift. The authors are grateful to Professor Karen Bieback (Heidelberg University; German Red Cross Blood Donor Service Baden-Württemberg–Hessen, Mannheim, Germany) for kindly providing MSCs and Birgit Huber (Soft Matter Synthesis Lab, KIT) for the synthesis of PEGDA.

PY - 2022/7

Y1 - 2022/7

N2 - Leukemia patients undergo chemotherapy to combat the leukemic cells (LCs) in the bone marrow. During therapy not only the LCs, but also the blood-producing hematopoietic stem and progenitor cells (HSPCs) may be destroyed. Chemotherapeutics targeting only the LCs are urgently needed to overcome this problem and minimize life-threatening side-effects. Predictive in vitro drug testing systems allowing simultaneous comparison of various experimental settings would enhance the efficiency of drug development. Here, we present a three-dimensional (3D) human leukemic bone marrow model perfused using a magnetic, parallelized culture system to ensure media exchange. Chemotherapeutic treatment of the acute myeloid leukemia cell line KG-1a in 3D magnetic hydrogels seeded with mesenchymal stem/stromal cells (MSCs) revealed a greater resistance of KG-1a compared to 2D culture. In 3D tricultures with HSPCs, MSCs and KG-1a, imitating leukemic bone marrow, HSPC proliferation decreased while KG-1a cells remained unaffected post treatment. Non-invasive metabolic profiling enabled continuous monitoring of the system. Our results highlight the importance of using biomimetic 3D platforms with proper media exchange and co-cultures for creating in vivo-like conditions to enable in vitro drug testing. This system is a step towards drug testing in biomimetic, parallelized in vitro approaches, facilitating the discovery of new anti-leukemic drugs.

AB - Leukemia patients undergo chemotherapy to combat the leukemic cells (LCs) in the bone marrow. During therapy not only the LCs, but also the blood-producing hematopoietic stem and progenitor cells (HSPCs) may be destroyed. Chemotherapeutics targeting only the LCs are urgently needed to overcome this problem and minimize life-threatening side-effects. Predictive in vitro drug testing systems allowing simultaneous comparison of various experimental settings would enhance the efficiency of drug development. Here, we present a three-dimensional (3D) human leukemic bone marrow model perfused using a magnetic, parallelized culture system to ensure media exchange. Chemotherapeutic treatment of the acute myeloid leukemia cell line KG-1a in 3D magnetic hydrogels seeded with mesenchymal stem/stromal cells (MSCs) revealed a greater resistance of KG-1a compared to 2D culture. In 3D tricultures with HSPCs, MSCs and KG-1a, imitating leukemic bone marrow, HSPC proliferation decreased while KG-1a cells remained unaffected post treatment. Non-invasive metabolic profiling enabled continuous monitoring of the system. Our results highlight the importance of using biomimetic 3D platforms with proper media exchange and co-cultures for creating in vivo-like conditions to enable in vitro drug testing. This system is a step towards drug testing in biomimetic, parallelized in vitro approaches, facilitating the discovery of new anti-leukemic drugs.

KW - Chemotherapeutics

KW - Drug testing

KW - Hematopoietic stem and progenitor cells

KW - Leukemic niche

KW - Magnetic hydrogels

UR - http://www.scopus.com/inward/record.url?scp=85130645094&partnerID=8YFLogxK

U2 - 10.1088/1758-5090/ac6a7e

DO - 10.1088/1758-5090/ac6a7e

M3 - Article

C2 - 35472717

AN - SCOPUS:85130645094

VL - 14

JO - Biofabrication

JF - Biofabrication

SN - 1758-5082

IS - 3

M1 - 035011

ER -