Neuroinflammation, a complex immune response within the central nervous system, stands as both a shield and a sword in neurological health. It involves the activation of immune cells like microglia and astrocytes, the release of signaling molecules, and phagocytosis of pathogens. While it serves as a frontline defense against threats, dysregulation can fuel the progression of neurological disorders such as Alzheimer’s and Parkinson’s disease, as well as multiple sclerosis.
At Scantox Neuro, we are offering advanced in vitro models with high translational relevance to evaluate the efficacy of therapeutics across various cell systems, protocols, and assays.
As the field of induced pluripotent stem cells (iPSCs) advances, we are constantly improving protocols. Leveraging our expertise with immortalized cell lines and rodent primary cells, we now established lipopolysaccharide (LPS)-induced cytokine release and Aβ phagocytosis assays in iPSC-derived human microglia.
Incubating differentiated iPSC-derived human microglia with LPS for 24 h leads to a significantly increased secretion of relevant cytokines, which can be reversed by reference items like dexamethasone (Dexa; Figure 1), providing a fast and reproducible screening tool for anti-inflammatory drugs.
Figure 1: Increased cytokine release in LPS-stimulated iPSC-derived microglia can be reversed with dexamethasone treatment. After 24 h stimulation, supernatants were collected and analyzed for three cytokines IL-8, IL-6 and TNF-alpha. Data are shown as pg/mL supernatant. Mean+SEM, n=4-7 per group. One way ANOVA with Bonferroni post hoc test. *p<0.05; **p<0.01. Dexa, dexamethasone; LPS, lipopolysaccharide; VC, vehicle control.
The same cells can also be used to assess phagocytosis modulators. Addition of recombinant Aβ1-42 coupled to pH-sensitive pHrodo™ Red label to iPSC-derived microglial cells allows to monitor uptake and lysosomal degradation measurable as increasing red fluorescence in the IncuCyte® Livecell imaging system (Figure 2).
Figure 2: Assessment of Aβ1-42 phagocytosis in LPS-treated iPSC-derived human microglia. Aβ1-42 phagocytosis was measured for 20 h using pHrodo™ Red labelled Aβ1-42 and IncuCyte® Livecell imaging. n=6 per group. Mean ± SEM. Two-way ANOVA with Bonferroni’s post hoc test. Representative videos of LPS- and VC-treated iPSC-derived human microglia are shown on the right. Dexa, dexamethasone; LPS, lipopolysaccharide; VC, vehicle control.
In a collaborative project with the NIH, this method was already successfully used to evaluate the effect of hydroxychloroquine on Aβ1-42 clearance. (Varma et al., 2023).
Contact us today to get your in vitro study in iPSC-derived microglia started.
