Differentiation of hiPSCs into Microglia

WT/WT and ΔE9/ΔE9 hiPSCs were differentiated into MGLs using a previously described protocol (McQuade et al., 2018 (link)). Briefly, hiPSCs were differentiated into hematopoietic progenitor cells (HPCs) using the STEMdiff™ Hematopoietic Kit (#05310, Stem cell Technologies). HPCs were either frozen using Bambanker HRM freezing media (#BBH01; Bulldog Bio) or further differentiated into MGLs. HPCs were cultured in microglia differentiation medium comprised of DMEM/F12 (#11039047; Thermo Fisher Scientific), B27 (#17504-044; Thermo Fisher Scientific), N2 (#17502-048; Thermo Fisher Scientific), insulin-transferrin-selenite (#41400045; Thermo Fisher Scientific), non-essential amino acids (#11140050; Thermo Fisher Scientific), Glutamax (#35050061; Thermo Fisher Scientific), human insulin (#I2643-25 mg; Sigma) and monothioglycerol (#M1753; Sigma) supplemented with 25 ng/ml human M-CSF (#PHC9501; Thermo Fisher Scientific), 50 ng/ml TGF-β1 (#130-108-969; Miltenyl), and 100 ng/ml IL-34 (#200-34; Peprotech). After 24 days in this medium, two additional cytokines, 100 ng/ml CD200 (#C311; NovoProtein), and 100 ng/ml CX3CL1 (#300-31; PeproTech), were added to the medium described above to mature MGLs. MGLs were cultured in this new medium for an additional week and then harvested for experiments.

Free full text: Click here

Mishra S., Kinoshita C., Axtman A.D, & Young J.E. (2022). Evaluation of a Selective Chemical Probe Validates That CK2 Mediates Neuroinflammation in a Human Induced Pluripotent Stem Cell-Derived Mircroglial Model. Frontiers in Molecular Neuroscience, 15, 824956.

Publication 2022

STEMdiff™ Hematopoietic Kit DMEM/F12 insulin-transferrin-selenite non-essential amino acids Glutamax human insulin monothioglycerol IL-34 CD200 CX3CL1

Cd200 Cultured medium Cx3cl1 Cytokines Essential amino acids Hematopoietic Hematopoietic progenitor cells Hipscs Human Il 34 Insulin M csf Microglia Monothioglycerol Selenite Stem cell Tgf β1 Transferrin

Corresponding Organization : University of North Carolina at Chapel Hill

Top 5 similar protocols

Protocol cited in 2 other protocols

Variable analysis

independent variables

Genotype of hiPSCs (WT/WT and ΔE9/ΔE9)

dependent variables

Differentiation of hiPSCs into MGLs

control variables

Differentiation protocol used to generate hematopoietic progenitor cells (HPCs) from hiPSCs
Culture conditions for HPC differentiation into MGLs (DMEM/F12 medium, growth factors, and supplements)
Duration of HPC differentiation into MGLs (24 days + 1 week)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!