Single-cell RNA sequencing of intramedullary canal tissue to improve methods for studying fracture repair biology

The ability to study the bone microenvironment of failed fracture healing may lead to biomarkers for fracture nonunion. Herein the authors describe a technique for isolating individual cells suitable for single-cell RNA sequencing analyses from intramedullary canal tissue collected by reaming during...

Full description

Saved in:
Bibliographic Details
Main Authors: James M Dominguez, Sharon M Moe, Neal X Chen, Todd O McKinley, Krista M Brown, Yunlong Liu, Hongyu Gao, Roman M Natoli
Format: Article
Language:English
Published: Taylor & Francis Group 2021-08-01
Series:BioTechniques
Subjects:
Online Access:https://www.future-science.com/doi/10.2144/btn-2021-0002
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850152832962920448
author James M Dominguez
Sharon M Moe
Neal X Chen
Todd O McKinley
Krista M Brown
Yunlong Liu
Hongyu Gao
Roman M Natoli
author_facet James M Dominguez
Sharon M Moe
Neal X Chen
Todd O McKinley
Krista M Brown
Yunlong Liu
Hongyu Gao
Roman M Natoli
author_sort James M Dominguez
collection DOAJ
description The ability to study the bone microenvironment of failed fracture healing may lead to biomarkers for fracture nonunion. Herein the authors describe a technique for isolating individual cells suitable for single-cell RNA sequencing analyses from intramedullary canal tissue collected by reaming during surgery. The purpose was to detail challenges and solutions inherent to the collection and processing of intramedullary canal tissue samples. The authors then examined single-cell RNA sequencing data from fresh and reanimated samples to demonstrate the feasibility of this approach for prospective studies.
format Article
id doaj-art-cdd5b915484a40638d1700d57d37cbf7
institution OA Journals
issn 0736-6205
1940-9818
language English
publishDate 2021-08-01
publisher Taylor & Francis Group
record_format Article
series BioTechniques
spelling doaj-art-cdd5b915484a40638d1700d57d37cbf72025-08-20T02:25:51ZengTaylor & Francis GroupBioTechniques0736-62051940-98182021-08-0171243143810.2144/btn-2021-0002Single-cell RNA sequencing of intramedullary canal tissue to improve methods for studying fracture repair biologyJames M Dominguez0Sharon M Moe1Neal X Chen2Todd O McKinley3Krista M Brown4Yunlong Liu5Hongyu Gao6Roman M Natoli71Department of Medicine, Division of Nephrology, Indiana University School of Medicine, Indianapolis, IN 46202, USA1Department of Medicine, Division of Nephrology, Indiana University School of Medicine, Indianapolis, IN 46202, USA1Department of Medicine, Division of Nephrology, Indiana University School of Medicine, Indianapolis, IN 46202, USA2Department of Orthopaedic Surgery, Indiana University Health Methodist Hospital, Indianapolis, IN 46202, USA2Department of Orthopaedic Surgery, Indiana University Health Methodist Hospital, Indianapolis, IN 46202, USA3Department of Medical & Molecular Genetics, Center for Medical Genomics, Indiana University School of Medicine, Indianapolis, IN 46202, USA3Department of Medical & Molecular Genetics, Center for Medical Genomics, Indiana University School of Medicine, Indianapolis, IN 46202, USA2Department of Orthopaedic Surgery, Indiana University Health Methodist Hospital, Indianapolis, IN 46202, USAThe ability to study the bone microenvironment of failed fracture healing may lead to biomarkers for fracture nonunion. Herein the authors describe a technique for isolating individual cells suitable for single-cell RNA sequencing analyses from intramedullary canal tissue collected by reaming during surgery. The purpose was to detail challenges and solutions inherent to the collection and processing of intramedullary canal tissue samples. The authors then examined single-cell RNA sequencing data from fresh and reanimated samples to demonstrate the feasibility of this approach for prospective studies.https://www.future-science.com/doi/10.2144/btn-2021-0002cell isolationcryopreservationfracturenonunionsingle-cell RNA sequencing
spellingShingle James M Dominguez
Sharon M Moe
Neal X Chen
Todd O McKinley
Krista M Brown
Yunlong Liu
Hongyu Gao
Roman M Natoli
Single-cell RNA sequencing of intramedullary canal tissue to improve methods for studying fracture repair biology
BioTechniques
cell isolation
cryopreservation
fracture
nonunion
single-cell RNA sequencing
title Single-cell RNA sequencing of intramedullary canal tissue to improve methods for studying fracture repair biology
title_full Single-cell RNA sequencing of intramedullary canal tissue to improve methods for studying fracture repair biology
title_fullStr Single-cell RNA sequencing of intramedullary canal tissue to improve methods for studying fracture repair biology
title_full_unstemmed Single-cell RNA sequencing of intramedullary canal tissue to improve methods for studying fracture repair biology
title_short Single-cell RNA sequencing of intramedullary canal tissue to improve methods for studying fracture repair biology
title_sort single cell rna sequencing of intramedullary canal tissue to improve methods for studying fracture repair biology
topic cell isolation
cryopreservation
fracture
nonunion
single-cell RNA sequencing
url https://www.future-science.com/doi/10.2144/btn-2021-0002
work_keys_str_mv AT jamesmdominguez singlecellrnasequencingofintramedullarycanaltissuetoimprovemethodsforstudyingfracturerepairbiology
AT sharonmmoe singlecellrnasequencingofintramedullarycanaltissuetoimprovemethodsforstudyingfracturerepairbiology
AT nealxchen singlecellrnasequencingofintramedullarycanaltissuetoimprovemethodsforstudyingfracturerepairbiology
AT toddomckinley singlecellrnasequencingofintramedullarycanaltissuetoimprovemethodsforstudyingfracturerepairbiology
AT kristambrown singlecellrnasequencingofintramedullarycanaltissuetoimprovemethodsforstudyingfracturerepairbiology
AT yunlongliu singlecellrnasequencingofintramedullarycanaltissuetoimprovemethodsforstudyingfracturerepairbiology
AT hongyugao singlecellrnasequencingofintramedullarycanaltissuetoimprovemethodsforstudyingfracturerepairbiology
AT romanmnatoli singlecellrnasequencingofintramedullarycanaltissuetoimprovemethodsforstudyingfracturerepairbiology