A new non-enzymatic method for isolating human intervertebral disc cells preserves the phenotype of nucleus pulposus cells.

dc.contributor.author

Tang, Xinyan

dc.contributor.author

Richardson, William J

dc.contributor.author

Fitch, Robert D

dc.contributor.author

Brown, Christopher R

dc.contributor.author

Isaacs, Robert E

dc.contributor.author

Chen, Jun

dc.date.accessioned

2024-08-15T13:10:53Z

dc.date.available

2024-08-15T13:10:53Z

dc.date.issued

2014-12

dc.description.abstract

Cells isolated from intervertebral disc (IVD) tissues of human surgical samples are one of potential sources for the IVD cellular therapy. The purpose of this study was to develop a new non-enzymatic method, "tissue incubation", for isolating human IVD cells. The IVD tissues of annulus fibrosus (AF) and nucleus pulposus (NP) were incubated separately in tissue culture flasks with culture medium. After 7-10 days incubation, cells were able to migrate out of IVD tissues and proliferate in vitro. After 3-4 weeks culture, expanded cells were harvested by trypsinization, and the remaining tissues were transferred to a new flask for another round of incubation. The molecular phenotype of IVD cells from juvenile and adult human samples was evaluated by both flow cytometry analysis and immunocytochemical staining for the expression of protein markers of NP cells (CD24, CD54, CD239, integrin α6 and laminin α5). Flow cytometry confirmed that both AF and NP cells of all ages positively expressed CD54 and integrin α6, with higher expression levels in NP cells than in AF cells for the juvenile group sample. However, CD24 expression was only found in juvenile NP cells, and not in AF or older disc cells. Similar expression patterns for NP markers were also confirmed by immunocytochemistry. In summary, this new non-enzymatic tissue incubation method for cell isolation preserves molecular phenotypic markers of NP cells and may provide a valuable cell source for the study of NP regeneration strategies.

dc.identifier.issn

0920-9069

dc.identifier.issn

1573-0778

dc.identifier.uri

https://hdl.handle.net/10161/31392

dc.language

eng

dc.publisher

Springer Science and Business Media LLC

dc.relation.ispartof

Cytotechnology

dc.relation.isversionof

10.1007/s10616-013-9650-7

dc.rights.uri

https://creativecommons.org/licenses/by-nc/4.0

dc.subject

Science & Technology

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Life Sciences & Biomedicine

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Biotechnology & Applied Microbiology

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Cell Biology

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Nucleus pulposus

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Tissue culture

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Phenotype

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Intervertebral disc

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Integrin

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MESENCHYMAL-STEM-CELLS

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GENE-EXPRESSION

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IN-VITRO

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PROGENITOR CELLS

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DEGENERATION

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MATRIX

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MODEL

dc.subject

TRANSPLANTATION

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REGENERATION

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DIFFERENTIATION

dc.title

A new non-enzymatic method for isolating human intervertebral disc cells preserves the phenotype of nucleus pulposus cells.

dc.type

Journal article

duke.contributor.orcid

Richardson, William J|0000-0001-9608-199X|0000-0002-8750-7263|0009-0003-7526-7797

pubs.begin-page

979

pubs.end-page

986

pubs.issue

6

pubs.organisational-group

Duke

pubs.organisational-group

School of Medicine

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Staff

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Clinical Science Departments

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Orthopaedic Surgery

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Pediatrics

pubs.organisational-group

University Initiatives & Academic Support Units

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University Institutes and Centers

pubs.organisational-group

Duke Global Health Institute

pubs.organisational-group

Initiatives

pubs.organisational-group

Neurosurgery

pubs.organisational-group

Duke Innovation & Entrepreneurship

pubs.publication-status

Published

pubs.volume

66

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