"iMacroMAT": iPSC-derived macrophages for customization of Pyrogen Testing

Scalable and adaptable production of macrophages for testing parenteral substances in the Monocyte Activation Test to regulatory compliance

Current efforts by the EDQM (European Directorate for the Quality of Medicines) and the FDA's (Food and Drug Administration) Modernization Act 2.0 aim to eliminate the use of animals or animal-derived materials for drug safety testing and mandate the development and use of alternative testing methods. Specifically, the Rabbit Pyrogen Test will soon be excluded from the Pharmacopoeias worldwide.

Immune cells in the human body, particularly monocytes/macrophages, are essential in the early detection of pathogens and mediating inflammatory responses. These key functions can be industrially integrated into the development and testing of drugs, e.g. in the Monocyte Activation Test (MAT). Due to various disadvantages of primary human cells such as availability, standardizable, and genetic background, they are insufficient for the safety assessment of parenteral drugs, and cell lines are often unsuitable for safety testing purposes due to their non-physiological properties.

To address these limitations, we offer the characterization and scaling of designer immune cells derived from induced pluripotent stem cells (iPSC), providing a reliable and efficient alternative for drug safety testing according to individual needs.

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Our approach: Taken the flexibility of the iPSC system to customize macrophages

"iMacroMAT"

Testing of drugs, active ingredients, molecules, and proteins for potential contaminants using the "iMacroMAT" - applying human macrophages derived from induced pluripotent stem cells (iPSCs) as a cellular source for the Monocyte Activation Test (MAT).

Our iPSC-based approach includes:

iPSC lines from donors of different genders, genetic backgrounds, and ethnicities
Disease-specific iPSC lines upon request

Our expertise and services

Based on our extensive expertise and unique know-how in iPSC-derived macrophages, we offer innovative solutions to enhance current MAT formats and develop next-generation techniques tailored to your needs.

We provide

Defined iPSC lines with access to material from the same donor
Stable and well-characterized genetic backgrounds of iPSCs and their macrophage derivatives
A ready-to-use macrophage manufacturing pipeline - scalable and flexible to meet your specific requirements
A macrophage-based cell product enabling entirely new MAT designs in terms of cell quantity and time of analysis
Fully animal-free manufacturing and MAT development
Development of off-the-shelf cryopreservation strategies for on-demand macrophage production
Genetic engineering approaches to modernize the MAT, improving handling and enabling readouts for automated safety assessment - MAT-on-a-Chip

Production and application of human macrophages from induced pluripotent stem cells (iPSCs)

The diagram describes the production of human macrophages from induced pluripotent stem cells (iPSCs) — © Fraunhofer ITEM, created with Biorender

New stem cell technologies such as induced pluripotent stem cells (iPSC), honored with the Nobel Prize in Physiology or Medicine in 2012), in combination with advanced bioprocess developments, have enabled the creation of donor-independent immune cell products (e.g. macrophages). Human macrophages can now be produced from iPSCs under controlled conditions and using scalable methods, matching the functional and physiological properties of cells in the human body.

This key technology for the standardized production of human macrophages allows their use in detecting a wide range of pyrogens from both non-endotoxin (bacteria, viruses, fungi, process contamination) and endotoxin (bacteria) origins. iPSC-macrophages can be universally adapted to new testing procedures and produced on a large scale in a cost-efficient manner.

New approaches in drug safety

Thus, iPSC-macrophages represent a new and more efficient tool for the MAT and its further development. Under the iMacroMAT technology, we offer iPSC-macrophages based testing and further development.

Vorteile der iMacroMAT-Technologie

Animal-free & human origin

Completely animal free cell product (from production to application, no FCS)
Reacts to all human-relevant pyrogens
No continuous donation needed - no ethical concerns

High standardization

Validated immune cell product for specific applications
No variability due to same donor background

High sensitivity & speed

More sensitive to all kind of pyrogens compared to current standards
Reacts much faster and more reliable, response time only 4-8 hours
Scalable production and upside potential for platform technology

iMacro-MAT in comparison to MAT run with conventional cells

Macrophages are the sensors in the MAT. Our iPSC-derived Macrophages "iMacros" are compared to monocyte-derived macrophages (MDM) and Macrophage cell line (MM6) (Abdin et al.)

The diagram shows that iMacros are morphologically and functionally comparable to standard monocytes — © Fraunhofer ITEM, Nico Lachmann
iMacros are morphologically and functionally comparable to standard monocytes

The diagram shows that iMacros show a higher sensitivity to endotoxin-based pyrogens at different concentrations. — © Fraunhofer ITEM, Nico Lachmann
iMacros show a higher sensitivity to endotoxin-based pyrogens

The diagram shows iMacros that show a higher sensitivity to non-endotoxin-based pyrogens at different concentrations — © Fraunhofer ITEM, Nico Lachmann
iMacros show a higher sensitivity to non-endotoxin-based pyrogens

iMacros show a faster response to endotoxin stimulation compared to standard monocytes — © Fraunhofer ITEM, Nico Lachmann
iMacros show a faster response to endotoxin stimulation

Line chart. — © Fraunhofer ITEM, Nico Lachmann
NF-κB reporter iMacros show a correlation between IL-6 secretion and luminescence after LPS stimulation

Relevant publications

Abdin SM, Mansel F, Hashtchin AR, Ackermann M, Hansen G, Becker B, Kick B, Pham N, Dietz H, Schaniel C, Martin U, Spreitzer I, Lachmann N. "Sensor macrophages derived from human induced pluripotent stem cells to assess pyrogenic contaminations in parenteral drugs". Biofabrication. 2024 May 17;16(3). doi: 10.1088/1758-5090/ad4744.

Desai O, Köster M, Kloos D, Lachmann N, Hauser H, Poortinga A, Wirth D. "Ultrasound-triggered drug release in vivo from antibubble-loaded macrophages". J Control Release. 2024 Dec 19:378:365-376. doi: 10.1016/j.jconrel.2024.12.007.

Jacobsen C, Plückebaum N, Ssebyatika G, Beyer S, Mendes-Monteiro L, Wang J, Kropp KA, González-Motos V, Steinbrück L, Ritter B, Rodríguez-González C, Böning H, Nikolouli E, Kinchington PR, Lachmann N, Depledge DP, Krey T, Viejo-Borbolla A. "Viral modulation of type II interferon increases T cell adhesion and virus spread". Nature Communications. 2024 Jun 22;15(1):5318. doi: 10.1038/s41467-024-49657-4.

Paasch D, Lachmann N, "CAR macrophages tuning the immune symphony of anti-cancer therapies". Cell Stem Cell. 2024 Jun 6;31(6):791-793. doi: 10.1016/j.stem.2024.05.006.

Abdin SM, Paasch D, Lachmann N. "CAR macrophages on a fast track to solid tumor therapy". Nat Immunol. 2024 Jan;25(1):11-12. doi: 10.1038/s41590-023-01696-7.

Abdin SM, Paasch D, Kloos A, Oliveira MC, Jang MS, Ackermann M, Stamopoulou A, Mroch PJ, Falk CS, von Kaisenberg CS, Schambach A, Heuser M, Moritz T, Hansen G, Morgan M, Lachmann N. "Scalable generation of functional human iPSC-derived CAR-macrophages that efficiently eradicate CD19-positive leukemia". J Immunother Cancer. 2023 Dec 22;11(12):e007705. doi: 10.1136/jitc-2023-007705.

Malainou C, Abdin SM, Lachmann N, Matt U, Herold S "Alveolar macrophages in tissue homeostasis, inflammation, and infection: evolving concepts of therapeutic targeting". J Clin Invest. 2023 Oct 2;133(19):e170501

Forde AJ, Kolter J, Zwicky P, Baasch S, Lohrmann F, Eckert M, Gres V, Lagies S, Gorka O, Rambold AS, Buescher JM, Kammerer B, Lachmann N, Prinz M, Groß O, Pearce EJ, Becher B, Henneke P., "Metabolic rewiring tunes dermal macrophages in staphylococcal skin infection". Science Immunol. 2023 Aug 18;8(86):eadg3517. doi: 10.1126/sciimmunol.adg3517. Epub 2023 Aug 11.

Ackermann M, Rafiei Hashtchin A, Manstein F, Carvalho Oliveira M, Kempf H, Zweigerdt R, Lachmann N. "Continuous human iPSC-macrophage mass production by suspension culture in stirred tank bioreactors". Nat Protocols. 2022 Feb;17(2):513-539. doi: 10.1038/s41596-021-00654-7

Ackermann M, Kempf H, Hetzel M, Hesse C, Hashtchin AR, Brinkert K, Schott JW, Haake K, Kühnel MP, Glage S, Figueiredo C, Jonigk D, Sewald K, Schambach A, Wronski S, Moritz T, Martin U, Zweigerdt R, Munder A, Lachmann N "Bioreactor-based mass production of human iPSC-derived macrophages enables immunotherapies against bacterial airway infections". Nature Communications. 2018 Nov 30;9(1):5088. doi: 10.1038/s41467-018-07570-7.