2025 Hyper Recent •CC0 1.0 Universal

This work is dedicated to the public domain. No rights reserved.

Access Preprint From Server
July 16th, 2025
Version: 2
Cardiff University
cell biology
biorxiv

Defining the Limits of hPSC Derived Models: hPSC-MSNs Fail to Recapitulate Authentic Striatal Identity

Bartley, O. J. M.Open in Google Scholar•Vinh, N.-N.Open in Google Scholar•Lelos, M.Open in Google Scholar•Williams, N. M.Open in Google Scholar•Precious, S. V.Open in Google Scholar•Rosser, A. E.Open in Google Scholar

Human pluripotent stem cells (hPSCs) are increasingly used to model human disease and as donor cells for regenerative medicine. However, the fidelity of hPSC-derived cell types remains a major concern, particularly when these cells are intended to replicate complex or region-specific subtypes, such as those required to explore and treat neurological diseases. Medium spiny neurons (MSNs), the principal projection neurons of the striatum, are one such target cell type relevant to disorders such as Huntington's disease. While protocols for generating hPSC-derived MSNs (hPSC-MSNs) exist, the extent to which these cells faithfully recapitulate their genuine counterparts is unclear. Here, we generated isogenic human induced pluripotent stem cells (hiPSCs) from striatal (LGE) and non-neural (fibroblast) fetal tissues, and differentiated them into MSN-like cells alongside a naive human embryonic stem cell (hESC) line. Using DNA methylation profiling and single-cell RNA sequencing, we systematically compared the epigenetic and transcriptional features of these hPSC-MSNs to authentic fetal MSNs. Our findings reveal persistent epigenetic signatures inherited from the tissue of origin, which influence differentiation outcomes. While LGE-derived hiPSCs retained elements of a striatal-biased methylome and yielded MSN-like cells with enhanced similarity to authentic MSNs, all hPSC-MSNs remained epigenetically and transcriptionally distinct from genuine MSNs and we identified clusters of hPSC-derived cells with aberrant or incomplete phenotypes. These results demonstrate that even isogenic hiPSC lines exhibit variable differentiation potential due to residual epigenetic memory and protocol compatibility. We highlight the need for refined protocols and rigorous benchmarking of hPSC-derived models, particularly for regionally specified neuronal subtypes. Our study underscores the complex relationship between epigenetic status, cell lineage, protocol adaptation, and differentiation outcome.

Similar Papers

biorxiv
Fri Jul 18 2025
Transcriptome-wide mRNP condensation precedes stress granuleformation and excludes new mRNAs
Stress-induced mRNP condensation is conserved across eukaryotes, resulting in stress granule formation under intense stresses, yet the mRNA composition and function of these condensates remain unclear. Exposure of ribosome-free mRNA following stress is thought to cause condensation and stress granule formation through mRNA-sequence-dependent interactions, leading to disproportionate condensation o...
Glauninger, H.
•
Bard, J. A. M.
•
Wong Hickernell, C. J.
•
Velez, K. M.
...•
Drummond, D. A.
biorxiv
Thu Jul 17 2025
Airway Epithelial SARS-CoV-2 Infectious and Repair Responses: Relationships to Age, Sex, and Post-COVID Pulmonary Syndromes
The long-term pulmonary sequelae of SARS-CoV-2 respiratory infections reflect infection severity, innate and adaptive immunity, and respiratory epithelial repair. This study investigated the acute and reparative responses as a function of age and sex in primary human bronchial epithelial (HBE) cultures over a 14-day post-SARS-CoV-2 infection (dpi) protocol. SARS-CoV-2 infection peaked at 3 dpi wit...
Dang, H.
•
Edwards, C. E.
•
Kato, T.
•
Reidel, B.
...•
Boucher, R. C.
biorxiv
Thu Jul 17 2025
Photostimulation Improves Maturation of Human Photoreceptors
The human retina contains photoreceptor cells that detect light and enable vision. The development of these cells involves a tightly regulated cascade of structural and molecular events, and their dysfunction leads to irreversible blindness in many retinal diseases. Human retinal organoids derived from stem cells have become powerful tools to model retinal development and disease, but they often r...
Celiker, C.
•
Hruba, E.
•
Kyriakou, K.
•
Dorgau, B.
...•
Barta, T.
biorxiv
Thu Jul 17 2025
LIM Domain Proteins link molecular and global tension by recognizing strained actin in adhesions
Mechanotransduction is fundamental to cell signaling and depends on force-sensitive adhesion proteins. How these proteins differentiate and integrate their responses to tension remains an open question. We show mechanosensitive LIM domain proteins like zyxin detect global adhesion tension by recognizing strained actin within these structures. In sharp contrast, vinculin localization and intramolec...
Sala, S.
•
Chandrasekar, S.
•
Troughton, L.
•
Wu, H.
...•
Oakes, P. W.
biorxiv
Thu Jul 17 2025
Spef1 is a microvillar component that limits apical actomyosin contractility and preserves intestinal barrier function
Epithelial sheet integrity is established by adherent contacts that form between cells, at the interface between their apical and basolateral domains. Although cell contacts are reinforced by actomyosin contractility, which generates tension that propagates across the apical surface. How epithelial cells tune tension to reinforce junctions without compromising their physical barrier properties rem...
Tapia Pastrana, R.
•
Morales Obregon, E. A.
•
Hecht, G.
•
Tyska, M. J.
biorxiv
Thu Jul 17 2025
Long-term daily light exposure boosts photoreceptor maturation in retinal organoids
Induced pluripotent stem cell-derived retinal organoids (ROs) have become promising personalized models to study inherited retinal diseases (IRDs) and develop new innovative therapies. Although ROs mimic key retinal features and show some light responsiveness, their differentiation and maturation remain limited and lengthy. We hypothesize that standard dark culture conditions limit the expression ...
van Oosten, E. M.
•
Hoogendoorn, A. D. M.
•
Kieboom, W.
•
Berendsen, S. L.
...•
Garanto, A.
biorxiv
Thu Jul 17 2025
Host iron deficiency protects against Plasmodium infection and drives parasite molecular reprofiling.
Background: Iron deficiency, anemia and Plasmodium infection represent significant global health challenges with overlapping geographical distributions, particularly affecting pregnant women in Africa. Previous evidence suggests complex interactions between iron status and malaria susceptibility. However, the mechanisms and clinical implications of this relationship remain poorly understood. Metho...
Clucas, D.
•
Bennett, C.
•
Harding, R.
•
Pettikiriarachchi, A.
...•
Ataide, R.
biorxiv
Wed Jul 16 2025
VapA/Scs2 sustains polarized growth in Aspergillus nidulans by maintaining AP-2-mediated apical endocytosis
Growth of filamentous fungi is highly polarized requiring the coordinated apical delivery of cell wall components and plasma membrane (PM) material, primarily lipids and proteins, to hyphal tips via conventional vesicular secretion. Fungal growth also requires the tight coordination of exocytosis (secretion) with endocytosis and recycling of proteins and lipids, which occurs in a defined region be...
Georgiou, X.
•
Politi, S.
•
Amillis, S.
•
Diallinas, G.
biorxiv
Wed Jul 16 2025
PIKfyve governs endoplasmic reticulum-lysosome contacts to modulate endoplasmic reticulum dynamics
Lysosomes are essential to clear unwanted cellular material delivered by constant membrane fusion. Membrane fission is thus required to balance lysosome size, number, and composition. PIKfyve is a lipid kinase that converts phosphatidylinositol-3-phosphate [PtdIns(3)P] to phosphatidylinositol-3,5-bisphosphate [PtdIns(3,5)P2] and promotes lysosome fission since lysosomes coalesce into larger, but f...
Jenkins, N.
•
Adamji, Z.
•
Almasri, N. R.
•
Lomiyev, S.
•
Botelho, R. J.