A step‐by‐step protocol for isolation of murine nucleus pulposus cells
Abstract The intervertebral disc (IVD) is composed of three separate tissues with distinct origins and properties. Elucidating changes occurring in these tissues in response to injury or age is paramount to identify new therapies to better manage disc and spine degenerative conditions, including low...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2019-12-01
|
| Series: | JOR Spine |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jsp2.1073 |
| _version_ | 1811344609938243584 |
|---|---|
| author | Andrew Bratsman Greig Couasnay Florent Elefteriou |
| author_facet | Andrew Bratsman Greig Couasnay Florent Elefteriou |
| author_sort | Andrew Bratsman |
| collection | DOAJ |
| description | Abstract The intervertebral disc (IVD) is composed of three separate tissues with distinct origins and properties. Elucidating changes occurring in these tissues in response to injury or age is paramount to identify new therapies to better manage disc and spine degenerative conditions, including low back pain. Despite their small size and different mechanical load pattern compared to higher species, the use of mouse models represents a cost‐effective and powerful approach to better understand the formation, maintenance, and degeneration of the IVD. However, the isolation of the different compartments of the IVD is complicated by their diminutive size. Here, we describe a simple, step‐by‐step protocol for the isolation of the nucleus pulposus (NP) tissues that can then be processed for further analyses. Analysis from mouse NP tissues shows sufficient quantities of RNAs, purity of the NP fraction, and overall RNA quality for gene expression studies, and reveals no increase in expression of disc degeneration markers, including TNFa, IL1b, and Mmp1 up to 15 months of age in C57BL6 wildtype mice. |
| first_indexed | 2024-04-13T19:49:45Z |
| format | Article |
| id | doaj.art-14bc9452370d4c06a9b1f178a62a16b9 |
| institution | Directory Open Access Journal |
| issn | 2572-1143 |
| language | English |
| last_indexed | 2024-04-13T19:49:45Z |
| publishDate | 2019-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | JOR Spine |
| spelling | doaj.art-14bc9452370d4c06a9b1f178a62a16b92022-12-22T02:32:34ZengWileyJOR Spine2572-11432019-12-0124n/an/a10.1002/jsp2.1073A step‐by‐step protocol for isolation of murine nucleus pulposus cellsAndrew Bratsman0Greig Couasnay1Florent Elefteriou2Department of Orthopedic Surgery Baylor College of Medicine Houston TexasDepartment of Orthopedic Surgery Baylor College of Medicine Houston TexasDepartment of Orthopedic Surgery Baylor College of Medicine Houston TexasAbstract The intervertebral disc (IVD) is composed of three separate tissues with distinct origins and properties. Elucidating changes occurring in these tissues in response to injury or age is paramount to identify new therapies to better manage disc and spine degenerative conditions, including low back pain. Despite their small size and different mechanical load pattern compared to higher species, the use of mouse models represents a cost‐effective and powerful approach to better understand the formation, maintenance, and degeneration of the IVD. However, the isolation of the different compartments of the IVD is complicated by their diminutive size. Here, we describe a simple, step‐by‐step protocol for the isolation of the nucleus pulposus (NP) tissues that can then be processed for further analyses. Analysis from mouse NP tissues shows sufficient quantities of RNAs, purity of the NP fraction, and overall RNA quality for gene expression studies, and reveals no increase in expression of disc degeneration markers, including TNFa, IL1b, and Mmp1 up to 15 months of age in C57BL6 wildtype mice.https://doi.org/10.1002/jsp2.1073aginggene expressionintervertebral disc degenerationnucleus pulposus |
| spellingShingle | Andrew Bratsman Greig Couasnay Florent Elefteriou A step‐by‐step protocol for isolation of murine nucleus pulposus cells JOR Spine aging gene expression intervertebral disc degeneration nucleus pulposus |
| title | A step‐by‐step protocol for isolation of murine nucleus pulposus cells |
| title_full | A step‐by‐step protocol for isolation of murine nucleus pulposus cells |
| title_fullStr | A step‐by‐step protocol for isolation of murine nucleus pulposus cells |
| title_full_unstemmed | A step‐by‐step protocol for isolation of murine nucleus pulposus cells |
| title_short | A step‐by‐step protocol for isolation of murine nucleus pulposus cells |
| title_sort | step by step protocol for isolation of murine nucleus pulposus cells |
| topic | aging gene expression intervertebral disc degeneration nucleus pulposus |
| url | https://doi.org/10.1002/jsp2.1073 |
| work_keys_str_mv | AT andrewbratsman astepbystepprotocolforisolationofmurinenucleuspulposuscells AT greigcouasnay astepbystepprotocolforisolationofmurinenucleuspulposuscells AT florentelefteriou astepbystepprotocolforisolationofmurinenucleuspulposuscells AT andrewbratsman stepbystepprotocolforisolationofmurinenucleuspulposuscells AT greigcouasnay stepbystepprotocolforisolationofmurinenucleuspulposuscells AT florentelefteriou stepbystepprotocolforisolationofmurinenucleuspulposuscells |