Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells

Lanfer J, Winner B, Kaindl J, Krumm L, Acera MG, Neurath M, Regensburger M, Krach F (2022)

Publication Language: English

Publication Type: Journal article, Review article

Publication year: 2022

Journal

International Journal of Molecular Sciences MDPI

Book Volume: 23

Article Number: 4526

Journal Issue: 9

URI: https://www.mdpi.com/1422-0067/23/9/4526

DOI: 10.3390/ijms23094526

Abstract

Current protocols converting human induced pluripotent stem cells (iPSCs) into induced microglia-like cells (iMGL) are either dependent on overexpression of transcription factors or require substantial experience in stem-cell technologies. Here, we developed an easy-to-use two-step protocol to convert iPSCs into functional iMGL via: (1) highly efficient differentiation of hematopoietic progenitor cells (HPCs) from iPSCs, and (2) optimized maturation of HPCs to iMGL. A sequential harvesting approach led to an increased HPC yield. The protocol implemented a freezing step, thus allowing HPC biobanking and flexible timing of differentiation into iMGL. Our iMGL responded adequately to the inflammatory stimuli LPS, and iMGL RNAseq analysis matched those of other frequently used protocols. Comparing three different coating modalities, we increased the iMGL yield by culturing on uncoated glass surfaces, thereby retaining differentiation efficiency and functional hallmarks of iMGL. In summary, we provide a high-quality, easy-to-use protocol, rendering generation and functional studies on iMGL an accessible lab resource.

Authors with CRIS profile

Beate Winner Professur für Stammzell-Modelle seltener neuraler Erkrankungen Johanna Kaindl Professur für Molekulare Neurologie Laura Krumm Professur für Molekulare Neurologie Markus Neurath Lehrstuhl für Innere Medizin I Martin Regensburger Interdisziplinäres Zentrum für Klinische Forschung IZKF-Förderlinien

Additional Organisation(s)

Interdisziplinäres Zentrum für Klinische Forschung

Related research project(s)

Bavarian Research Association Interaction of Human Brain Cells (ForInter) March 1, 2019 - Dec. 31, 2023 E030: Th17 and beyond: immune-mediated neurotoxicity determines onset and progression in Parkinson’s disease April 1, 2020 - March 31, 2023 E030: Th17 and beyond: immune-mediated neurotoxicity determines onset and progression in Parkinson’s disease April 1, 2020 - March 31, 2023 N2: Sensorbasiertes Monitoring der Gangstörung bei HSP-Patienten mit Umsetzung im AlltagN7: Identifikation der Veränderungen im GSK3-Signalweg bei autosomal-rezessiven Typen der kompliziertenHSP anhand von neuralen Modellen aus induziert-pluripotenten Stammzellen (TreatHSP) N2: Sensorbasiertes Monitoring der Gangstörung bei HSP-Patienten mit Umsetzung im AlltagN7: Identifikation der Veränderungen im GSK3-Signalweg bei autosomal-rezessiven Typen der kompliziertenHSP anhand von neuralen Modellen aus induziert-pluripotenten Stammzellen May 1, 2019 - Jan. 31, 2023

Involved external institutions

University of California, San Diego US United States (USA) (US)

How to cite

APA:

Lanfer, J., Winner, B., Kaindl, J., Krumm, L., Acera, M.G., Neurath, M.,... Krach, F. (2022). Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells. International Journal of Molecular Sciences, 23(9). https://dx.doi.org/10.3390/ijms23094526

MLA:

Lanfer, Jonas, et al. "Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells." International Journal of Molecular Sciences 23.9 (2022).

BibTeX: Download