Astrocytes exhibit dynamic behavior in multiple sclerosis (MS), assuming both pro-inflammatory and anti-inflammatory phenotypes, yet their precise role in modelling the neuroinflammatory response in MS remains elusive. While mice are commonly employed for MS research, murine astrocytes fail to fully replicate the structural and functional complexity of their human counterparts, limiting the translation of findings from in vitro studies to the human context. Leveraging the unique characteristics of patient-derived iPSC astrocytes, this study aims to establish a platform for in vitro exploration of human neuron-astrocyte interactions within the structured environment of the mouse hippocampus by engrafting human iPSC-derived neurons and astrocytes into murine hippocampal slices. To create a better microenvironment to promote the growth of human cells, we will first deplete murine neurons and astrocytes from hippocampal mouse slices. We achieved the depletion of murine astrocyte via 4-hydroxytamoxifen (4-OHT) treatment in bpA x B6.Aldh1L1-Cre-ERT2 mouse slices, while we are currently testing the application of NMDA to induce excitotoxicity to deplete neurons. Following the successful depletion of murine astrocytes and neurons, we will engraft their human counterparts into the slices and use immunostaining and confocal imaging to verify the success of this procedure. Preliminary data of engrafted iPSC-derived astrocytes into hippocampal mouse slices with pre-hoc murine glial depletion demonstrated the feasibility of the development of humanized mouse slices (Fig. 1). The success of this project will significantly advance disease modelling using patient-derived cells in a more ecological in in vitro context, allowing to enhance our understanding of neuron-astrocyte interactions in MS.