Resolving the bone – optimizing decalcification in spatial transcriptomics and molecular pathology DOI
Shuoshuo Wang

Journal of Histotechnology, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 10

Published: Dec. 26, 2024

Bone tissue poses critical roadblocks for spatial transcriptomics and molecular pathology due to a combination of its dense, calcified matrix inadequate preservation biomolecules in conventional decalcification. Decalcification is complex nuanced histological process concomitantly preserve nucleic acids, proteins, architecture, ensuring integrity downstream assays. However, commonly used agents like formic hydrochloric while efficient, can degrade varying extents, complicating assays such as PCR, sequencing, immunohistochemistry, situ hybridization. Advances transcriptomics, both sequencing- imaging-based, emphasize the importance optimizing decalcification protocols improve research outcomes. This synoptic perspective article explores traditional modern methods, offering recommendations on technical methodological refinements achieving molecularly robust processing bone tissues pathology.

Language: Английский

Single-cell transcriptional profiling reveals a novel RAB13+ endothelial subpopulation and profibrotic mesenchymal cells in the aged human bone marrow DOI Open Access
Itziar Cenzano, Miguel Cócera,

A. Perez

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 29, 2025

ABSTRACT The bone marrow (BM) microenvironment plays a crucial role in regulating hematopoiesis, yet the molecular and functional changes associated with aging humans remain poorly understood. Using single-cell RNA sequencing (scRNA-seq), we uncovered transcriptional shifts BM endothelial cells (EC) mesenchymal stem (MSC) during aging. Our analysis revealed that aged sinusoidal EC adopt prothrombotic, exhibit mitochondrial dysfunction, have compromised vascular function. Additionally, identified unique arterial subset, present only individuals, elongation senescence processes characterized by RAB13 expression. MSC from subjects displayed an impaired matrix remodeling epithelial-mesenchymal transition, driven partly subpopulation of THY1 + profibrotic stromal absent young subjects. Aged were also increased ATP-oxidative metabolism reduced protein folding capacity. Finally, using immunofluorescent imaging spatial transcriptomics, confirmed presence senescent samples significant age-related cell-cell communication within niche. In summary, this work provides comprehensive view diversity, cellular interactions, organization MSC, offering novel insights potential targets could be exploited for preventing aged-associated humans.

Language: Английский

Citations

0
Resolving the bone – optimizing decalcification in spatial transcriptomics and molecular pathology DOI
Shuoshuo Wang

Journal of Histotechnology, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 10

Published: Dec. 26, 2024

Bone tissue poses critical roadblocks for spatial transcriptomics and molecular pathology due to a combination of its dense, calcified matrix inadequate preservation biomolecules in conventional decalcification. Decalcification is complex nuanced histological process concomitantly preserve nucleic acids, proteins, architecture, ensuring integrity downstream assays. However, commonly used agents like formic hydrochloric while efficient, can degrade varying extents, complicating assays such as PCR, sequencing, immunohistochemistry, situ hybridization. Advances transcriptomics, both sequencing- imaging-based, emphasize the importance optimizing decalcification protocols improve research outcomes. This synoptic perspective article explores traditional modern methods, offering recommendations on technical methodological refinements achieving molecularly robust processing bone tissues pathology.

Language: Английский

Citations

1