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

Area of Science:

• Stem cell biology
• Neuroscience
• Immunology

Background:

• Current methods for generating induced microglia-like cells (iMGL) from human induced pluripotent stem cells (iPSCs) are complex, requiring transcription factor manipulation or extensive expertise.
• There is a need for a simplified, accessible protocol for iMGL generation for research applications.

Purpose of the Study:

• To develop an easy-to-use, two-step protocol for converting iPSCs into functional iMGL.
• To optimize the differentiation and maturation process for increased yield and efficiency.
• To provide a reliable iMGL model for studying microglia function.

Main Methods:

• A two-step protocol involving differentiation of iPSCs into hematopoietic progenitor cells (HPCs) followed by optimized maturation into iMGL.
• Implementation of a sequential harvesting approach for increased HPC yield.
• Utilizing a freezing step for HPC biobanking and flexible iMGL differentiation timing.
• Comparison of different coating modalities, identifying uncoated glass surfaces as optimal for iMGL culture.

Main Results:

• The protocol successfully generated functional iMGL from iPSCs.
• A sequential harvesting approach enhanced HPC yield.
• Culturing on uncoated glass surfaces increased iMGL yield while maintaining differentiation efficiency and functional characteristics.
• Generated iMGL exhibited adequate responses to lipopolysaccharide (LPS) stimulation and comparable RNA sequencing profiles to existing protocols.

Conclusions:

• A high-quality, user-friendly protocol for generating functional iMGL from iPSCs has been established.
• This simplified method makes iMGL generation and functional studies more accessible for laboratory research.
• The protocol offers a valuable resource for advancing research in neuroinflammation and microglia biology.