SOP for snap-freezing tissues v1

Mimpen, Jolet Y; Paul, Claudia; Ramos‐Mucci, Lorenzo; Network, Tendon Seed; Cribbs, Adam P.; Baldwin, Mathew; Snelling, Sarah

doi:10.17504/protocols.io.b2tqqemw

Cited by 2 publications

(3 citation statements)

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“…The age, gender, affected side, and body mass index of the donors were collected (Table 1). Tendon was handled as previously described 15 . In short, tissue was collected in cold DMEM‐F12 media (Gibco) supplemented with 10% Fetal Bovine Serum (Labtech International) and 1% Penicillin/Streptomycin (Gibco) and transferred to the research facility for processing.…”

Section: Methodsmentioning

confidence: 99%

“…Tendon was handled as previously described. 15 In short, tissue was collected in cold DMEM-F12 media (Gibco) supplemented with 10% Fetal Bovine Serum (Labtech International) and 1% Penicillin/Streptomycin (Gibco) and transferred to the research facility for processing. Within 2 h of tissue collection, tendon was washed in PBS, fat and muscle were cut off, the tendon was cut into 1 cm pieces, and photographed to retain topographical reference.…”

Section: Tissue Acquisition and Processingmentioning

confidence: 99%

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Single nucleus and spatial transcriptomic profiling of healthy human hamstring tendon

Mimpen,

Ramos‐Mucci,

Paul

et al. 2024

The FASEB Journal

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The molecular and cellular basis of health in human tendons remains poorly understood. Among human tendons, hamstring tendon has markedly low pathology and can provide a prototypic healthy tendon reference. The aim of this study was to determine the transcriptomes and location of all cell types in healthy hamstring tendon. Using single nucleus RNA sequencing, we profiled the transcriptomes of 10 533 nuclei from four healthy donors and identified 12 distinct cell types. We confirmed the presence of two fibroblast cell types, endothelial cells, mural cells, and immune cells, and identified cell types previously unreported in tendons, including different skeletal muscle cell types, satellite cells, adipocytes, and undefined nervous system cells. The location of these cell types within tendon was defined using spatial transcriptomics and imaging, and potential transcriptional networks and cell–cell interactions were analyzed. We demonstrate that fibroblasts have the highest number of potential cell–cell interactions in our dataset, are present throughout the tendon, and play an important role in the production and organization of extracellular matrix, thus confirming their role as key regulators of hamstring tendon homeostasis. Overall, our findings underscore the complexity of the cellular networks that underpin healthy human tendon function and the central role of fibroblasts as key regulators of hamstring tendon tissue homeostasis.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Tissue Acquisition and Processingmentioning

confidence: 99%

Single nucleus and spatial transcriptomic profiling of healthy human hamstring tendon

Mimpen,

Ramos‐Mucci,

Paul

et al. 2024

The FASEB Journal

Self Cite

View full text Add to dashboard Cite

show abstract

“…The age, gender, affected side, and body mass index were collected (Table 1). Tendon was handled as previously described [11]. Tissue was collected in cold DMEM-F12 media supplemented with 10% Foetal Bovine Serum and 1% Penicillin/Streptomycin and transferred to the research facility for processing.…”

Section: Methodsmentioning

confidence: 99%

Single nucleus and spatial transcriptomic profiling of human healthy hamstring tendon

Mimpen

Ramos‐Mucci

Paul

et al. 2022

Preprint

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The molecular and cellular basis of health in human tendons remains poorly understood. Amongst human tendons, the hamstrings are the least likely to be injured or degenerate, providing a prototypic healthy tendon reference. The aim of this study was to define the transcriptome and location of all cell types in healthy hamstring tendon. We profiled the transcriptomes of 10,533 nuclei from 4 healthy donors using single-nucleus RNA sequencing (snRNA-seq) and identified 12 distinct cell types. We confirmed the presence of two fibroblast cell types, endothelial cells, mural cells, and immune cells, and revealed the presence of cell types previously unreported for tendon sites, including different skeletal muscle cell types, satellite cells, adipocytes, and nerve cells, which are undefined nervous system cells. Location of these cell types within tendon was defined using spatial transcriptomics and imaging, and transcriptional networks and cell-cell interactions were identified. We demonstrate that fibroblasts have a high number of potential cell-cell interactions, are present throughout the whole tendon tissue, and play an important role in the production and organisation of extracellular matrix, thus confirming their role as key regulators of hamstring tendon tissue homeostasis. Overall, our findings highlight the highly complex cellular networks underpinning tendon function and underpins the importance of fibroblasts as key regulators of hamstring tendon tissue homeostasis.

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SOP for snap-freezing tissues v1

Cited by 2 publications

References 0 publications

Single nucleus and spatial transcriptomic profiling of healthy human hamstring tendon

Single nucleus and spatial transcriptomic profiling of healthy human hamstring tendon

Single nucleus and spatial transcriptomic profiling of human healthy hamstring tendon

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