Hyperosmolar expansion medium improves nucleus pulposus cell phenotype.
cell‐based therapy
intervertebral disc
lower back pain
nucleus pulposus
osmolarity
regeneration
Journal
JOR spine
ISSN: 2572-1143
Titre abrégé: JOR Spine
Pays: United States
ID NLM: 101722350
Informations de publication
Date de publication:
Sep 2022
Sep 2022
Historique:
received:
16
04
2022
revised:
21
07
2022
accepted:
21
07
2022
entrez:
7
10
2022
pubmed:
8
10
2022
medline:
8
10
2022
Statut:
epublish
Résumé
Repopulating the degenerated intervertebral disc (IVD) with tissue-specific nucleus pulposus cells (NPCs) has already been shown to promote regeneration in various species. Yet the applicability of NPCs as cell-based therapy has been hampered by the low cell numbers that can be extracted from donor IVDs and their potentially limited regenerative capacity due to their degenerated phenotype. To optimize the expansion conditions, we investigated the effects of increasing culture medium osmolarity during expansion on the phenotype of dog NPCs and their ability to produce a healthy extracellular matrix (ECM) in a 3D culture model. Dog NPCs were expanded in expansion medium with a standard osmolarity of 300 mOsm/L or adjusted to 400 or 500 mOsm/L in both normoxic and hypoxic conditions. Following expansion, NPCs were cultured in a 3D culture model in chondrogenic culture medium with a standard osmolarity. Read-out parameters included cell proliferaton rate, morphology, phenotype and healthy ECM production. Increasing the expansion medium osmolarity from 300 to 500 mOsm/L resulted in NPCs with a more rounded morphology and a lower cell proliferation rate accompanied by the expression of several healthy NPC and progenitor markers at gene ( Altogether, our findings show that increasing medium osmolarity during expansion results in an NPC population with improved phenotype, which could enhance the potential of cell-based therapies for IVD regeneration.
Sections du résumé
Background
UNASSIGNED
Repopulating the degenerated intervertebral disc (IVD) with tissue-specific nucleus pulposus cells (NPCs) has already been shown to promote regeneration in various species. Yet the applicability of NPCs as cell-based therapy has been hampered by the low cell numbers that can be extracted from donor IVDs and their potentially limited regenerative capacity due to their degenerated phenotype. To optimize the expansion conditions, we investigated the effects of increasing culture medium osmolarity during expansion on the phenotype of dog NPCs and their ability to produce a healthy extracellular matrix (ECM) in a 3D culture model.
Methods
UNASSIGNED
Dog NPCs were expanded in expansion medium with a standard osmolarity of 300 mOsm/L or adjusted to 400 or 500 mOsm/L in both normoxic and hypoxic conditions. Following expansion, NPCs were cultured in a 3D culture model in chondrogenic culture medium with a standard osmolarity. Read-out parameters included cell proliferaton rate, morphology, phenotype and healthy ECM production.
Results
UNASSIGNED
Increasing the expansion medium osmolarity from 300 to 500 mOsm/L resulted in NPCs with a more rounded morphology and a lower cell proliferation rate accompanied by the expression of several healthy NPC and progenitor markers at gene (
Conclusions
UNASSIGNED
Altogether, our findings show that increasing medium osmolarity during expansion results in an NPC population with improved phenotype, which could enhance the potential of cell-based therapies for IVD regeneration.
Identifiants
pubmed: 36203869
doi: 10.1002/jsp2.1219
pii: JSP21219
pmc: PMC9520765
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e1219Informations de copyright
© 2022 The Authors. JOR Spine published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.
Déclaration de conflit d'intérêts
The authors declare no conflicts of interest.
Références
Spine (Phila Pa 1976). 2000 Feb 15;25(4):487-92
pubmed: 10707396
Tissue Eng Part C Methods. 2014 Aug;20(8):652-62
pubmed: 24304309
Eur Spine J. 2008 Dec;17 Suppl 4:452-8
pubmed: 19005704
Ann Transl Med. 2020 Mar;8(6):299
pubmed: 32355743
JOR Spine. 2019 Mar 01;2(1):e1043
pubmed: 31463457
Exp Hematol. 2002 Jan;30(1):42-8
pubmed: 11823036
Spine (Phila Pa 1976). 2012 Mar 1;37(5):351-8
pubmed: 21544011
JOR Spine. 2018 Dec;1(4):e1036
pubmed: 30895277
Rheumatology (Oxford). 2009 Jan;48(1):5-10
pubmed: 18854342
Biochem Biophys Res Commun. 2000 Apr 2;270(1):52-61
pubmed: 10733904
Oncotarget. 2018 May 29;9(41):26507-26526
pubmed: 29899873
Eur Cell Mater. 2015 Sep 21;30:132-46; discussion 146-7
pubmed: 26388616
J Spine Surg. 2019 Dec;5(4):561-583
pubmed: 32043007
Eur Spine J. 2008 Dec;17 Suppl 4:441-51
pubmed: 19005698
Biosci Rep. 2019 Sep 20;39(9):
pubmed: 31471533
J Orthop Res. 2022 Sep;40(9):2089-2102
pubmed: 34812520
Spine (Phila Pa 1976). 1999 Dec 1;24(23):2419-25
pubmed: 10626303
Front Cell Dev Biol. 2022 Mar 14;9:780749
pubmed: 35359916
Spine (Phila Pa 1976). 2006 Jun 15;31(14):1522-31
pubmed: 16778683
Eur Spine J. 2012 May;21 Suppl 1:S10-9
pubmed: 22395304
Spine J. 2013 Nov;13(11):1590-6
pubmed: 23800820
Arthritis Res Ther. 2010;12(3):R100
pubmed: 20492652
Spine (Phila Pa 1976). 2001 Aug 15;26(16):1747-51; discussion 1752
pubmed: 11493844
Stem Cells Int. 2015;2015:946031
pubmed: 26074979
Spine (Phila Pa 1976). 2018 Mar 1;43(5):307-315
pubmed: 25856264
JOR Spine. 2021 Nov 24;4(4):e1175
pubmed: 35005441
J Biomech Eng. 2005 Dec;127(7):1121-6
pubmed: 16502654
Crit Rev Eukaryot Gene Expr. 2014;24(3):193-204
pubmed: 25072146
Am J Physiol Renal Physiol. 2000 Jun;278(6):F1006-12
pubmed: 10836989
Nat Rev Rheumatol. 2015 Apr;11(4):243-56
pubmed: 25708497
Sci Rep. 2017 May 4;7(1):1501
pubmed: 28473691
Cell Physiol Biochem. 2016;39(6):2216-2226
pubmed: 27832635
Exp Cell Res. 1999 Jan 10;246(1):129-37
pubmed: 9882522
Eur Spine J. 2012 Aug;21 Suppl 6:S839-49
pubmed: 21874295
Eur Spine J. 2008 Dec;17 Suppl 4:492-503
pubmed: 19005697
JOR Spine. 2018 Aug 02;1(3):e1027
pubmed: 31463447
J Orthop Res. 1992 Sep;10(5):677-90
pubmed: 1380073
Stem Cell Res Ther. 2016 May 23;7(1):75
pubmed: 27216150
Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
pubmed: 12824337
Acta Biochim Biophys Sin (Shanghai). 2017 Jan;49(1):1-13
pubmed: 27864283
J Tissue Eng Regen Med. 2018 Mar;12(3):642-652
pubmed: 28544701
Nat Rev Genet. 2012 Mar 13;13(4):227-32
pubmed: 22411467
Arthritis Res Ther. 2005;7(4):R732-45
pubmed: 15987475
Eur Cell Mater. 2021 Jan 27;41:121-141
pubmed: 33528024
Nat Commun. 2012;3:1264
pubmed: 23232394
Spine (Phila Pa 1976). 2007 Mar 1;32(5):495-502
pubmed: 17334282
J Orthop Res. 2016 Aug;34(8):1327-40
pubmed: 26910849
Clin Orthop Relat Res. 2001 Aug;(389):94-101
pubmed: 11501830
Spine (Phila Pa 1976). 2012 Apr 1;37(7):E430-8
pubmed: 22466575
Am J Physiol Renal Physiol. 2005 Oct;289(4):F803-7
pubmed: 15900024
Nucleic Acids Res. 2001 May 1;29(9):e45
pubmed: 11328886
Blood. 2011 Jan 13;117(2):459-69
pubmed: 20952688
Connect Tissue Res. 2001;42(3):197-207
pubmed: 11913491
Stem Cells Dev. 2018 Feb 1;27(3):147-165
pubmed: 29241405
Spine (Phila Pa 1976). 1998 Jul 15;23(14):1531-8; discussion 1539
pubmed: 9682309
J Bone Miner Res. 2009 Jun;24(6):992-1001
pubmed: 19138132
Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
pubmed: 9254694
J Orthop Res. 2015 Mar;33(3):283-93
pubmed: 25411088
Biomol Eng. 2007 Feb;24(1):5-21
pubmed: 16963315
Lancet. 2018 Jun 9;391(10137):2302
pubmed: 29573869
Transplantation. 2003 Feb 15;75(3):389-97
pubmed: 12589164
Spine (Phila Pa 1976). 2003 Sep 1;28(17):1945-53; discussion 1953
pubmed: 12973139
Vet J. 2013 Mar;195(3):282-91
pubmed: 23177522
JOR Spine. 2018 Jun 27;1(2):e1018
pubmed: 31463445
DNA Repair (Amst). 2009 Aug 6;8(8):930-43
pubmed: 19535302
Tissue Eng Part C Methods. 2018 Apr;24(4):222-232
pubmed: 29457534