Investigation of stemness and multipotency of equine adipose-derived mesenchymal stem cells (ASCs) from different fat sources in comparison with lipoma.
Cell differentiation
Cell proliferation
Lipoma
Mesenchymal stem cells
Journal
Stem cell research & therapy
ISSN: 1757-6512
Titre abrégé: Stem Cell Res Ther
Pays: England
ID NLM: 101527581
Informations de publication
Date de publication:
22 10 2019
22 10 2019
Historique:
received:
17
04
2019
accepted:
26
09
2019
revised:
25
08
2019
entrez:
24
10
2019
pubmed:
24
10
2019
medline:
24
7
2020
Statut:
epublish
Résumé
Adipose tissue-derived mesenchymal stem cells (ASCs) offer a promising cell source for therapeutic applications in musculoskeletal disorders. The appropriate selection of ASCs from various fat depots for cell-based therapy is challenging. The present study aims to compare stemness and multipotency of ASCs derived from retroperitoneal (RP), subcutaneous (SC), and lipoma (LP) fat to assess their usefulness for clinical application. Equine ASCs from the three fat tissue sources were isolated and characterized. The cell viability, proliferation, and self-renewal were evaluated using MTT, sulforhodamine B, and colony forming unit (CFU) assays. Stem cell relative marker CD44, CD90, and CD105 and tumor marker CA9 and osteopontin (OPN) expression were quantified using RT-qPCR. Multipotency of ASCs for adipogenic, osteogenic, and chondrogenic differentiation was examined by quantifying Oil Red O and Alizarin Red S staining, alkaline phosphatase activity (ALP), and expression of differentiation relative markers. All data were statistically analyzed using ANOVA. RP fat-derived ASCs showed a higher cell proliferation rate compared to SC and LP derived cells. In contrast, ASCs from lipoma displayed a lower proliferation rate and impaired CFU capacities. The expression of CD44, CD90, and CD105 was upregulated in RP and SC derived cells but not in LP cells. RP fat-derived cells displayed a higher adipogenic potential compared to SC and LP cells. Although ASCs from all fat sources showed enhanced ALP activity following osteogenic differentiation, SC fat-derived cells revealed upregulated ALP and bone morphogenetic protein-2 expression together with a higher calcium deposition. We found an enhanced chondrogenic potency of RP and SC fat-derived cells as shown by Alcian blue staining and upregulation of aggrecan (Aggre), cartilage oligomeric matrix protein precursor (COMP), and collagen 2a1 (Col2a1) expression compared to LP. The expression of OPN and CA9 was exclusively upregulated in the ASCs of LP. The results provide evidence of variation in ASC performance not only between normal fat depots but also compared to LP cells which suggest a different molecular regulation controlling the cell fate. These data provided are useful when considering a source for cell replacement therapy in equine veterinary medicine.
Sections du résumé
BACKGROUND
Adipose tissue-derived mesenchymal stem cells (ASCs) offer a promising cell source for therapeutic applications in musculoskeletal disorders. The appropriate selection of ASCs from various fat depots for cell-based therapy is challenging. The present study aims to compare stemness and multipotency of ASCs derived from retroperitoneal (RP), subcutaneous (SC), and lipoma (LP) fat to assess their usefulness for clinical application.
METHODS
Equine ASCs from the three fat tissue sources were isolated and characterized. The cell viability, proliferation, and self-renewal were evaluated using MTT, sulforhodamine B, and colony forming unit (CFU) assays. Stem cell relative marker CD44, CD90, and CD105 and tumor marker CA9 and osteopontin (OPN) expression were quantified using RT-qPCR. Multipotency of ASCs for adipogenic, osteogenic, and chondrogenic differentiation was examined by quantifying Oil Red O and Alizarin Red S staining, alkaline phosphatase activity (ALP), and expression of differentiation relative markers. All data were statistically analyzed using ANOVA.
RESULTS
RP fat-derived ASCs showed a higher cell proliferation rate compared to SC and LP derived cells. In contrast, ASCs from lipoma displayed a lower proliferation rate and impaired CFU capacities. The expression of CD44, CD90, and CD105 was upregulated in RP and SC derived cells but not in LP cells. RP fat-derived cells displayed a higher adipogenic potential compared to SC and LP cells. Although ASCs from all fat sources showed enhanced ALP activity following osteogenic differentiation, SC fat-derived cells revealed upregulated ALP and bone morphogenetic protein-2 expression together with a higher calcium deposition. We found an enhanced chondrogenic potency of RP and SC fat-derived cells as shown by Alcian blue staining and upregulation of aggrecan (Aggre), cartilage oligomeric matrix protein precursor (COMP), and collagen 2a1 (Col2a1) expression compared to LP. The expression of OPN and CA9 was exclusively upregulated in the ASCs of LP.
CONCLUSIONS
The results provide evidence of variation in ASC performance not only between normal fat depots but also compared to LP cells which suggest a different molecular regulation controlling the cell fate. These data provided are useful when considering a source for cell replacement therapy in equine veterinary medicine.
Identifiants
pubmed: 31640774
doi: 10.1186/s13287-019-1429-0
pii: 10.1186/s13287-019-1429-0
pmc: PMC6805636
doi:
Types de publication
Comparative Study
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
309Références
Stem Cells Dev. 2008 Aug;17(4):761-73
pubmed: 18393634
Zhonghua Zheng Xing Wai Ke Za Zhi. 2010 Mar;26(2):125-32
pubmed: 20540318
Clin Cases Miner Bone Metab. 2017 May-Aug;14(2):217-226
pubmed: 29263737
Cell Commun Signal. 2011 May 14;9:12
pubmed: 21569606
Cytotherapy. 2006;8(4):315-7
pubmed: 16923606
Br J Haematol. 2005 Apr;129(1):118-29
pubmed: 15801964
Int J Biol Sci. 2015 Jul 18;11(10):1127-39
pubmed: 26327807
Clin Orthop Relat Res. 2017 Jun;475(6):1693-1701
pubmed: 28155209
Aesthetic Plast Surg. 2012 Oct;36(5):1164-7
pubmed: 22660950
Stem Cells. 2006 May;24(5):1294-301
pubmed: 16410387
Ann Plast Surg. 2008 May;60(5):538-44
pubmed: 18434829
J Craniofac Surg. 2013 Jul;24(4):1310-3
pubmed: 23851795
Cells. 2018 Dec 08;7(12):
pubmed: 30544806
Histochem Cell Biol. 2005 Aug;124(2):113-21
pubmed: 16032396
J Endocrinol. 2007 May;193(2):235-43
pubmed: 17470514
Trends Biotechnol. 2006 Apr;24(4):150-4
pubmed: 16488036
Adv Biochem Eng Biotechnol. 2010;123:219-63
pubmed: 20309674
Biochem Biophys Res Commun. 2007 Oct 5;361(4):883-9
pubmed: 17679141
Tissue Eng. 2001 Apr;7(2):211-28
pubmed: 11304456
Cell Transplant. 2015;24(11):2337-51
pubmed: 25562327
J Vet Sci. 2018 Jan 31;19(1):13-20
pubmed: 28693305
Can J Vet Res. 2016 Oct;80(4):294-301
pubmed: 27733784
J Cell Mol Med. 2017 Sep;21(9):2153-2162
pubmed: 28374574
Cytotherapy. 2013 Jun;15(6):641-8
pubmed: 23570660
Circ Res. 2007 May 11;100(9):1249-60
pubmed: 17495232
Vet Res Commun. 2011 Aug;35(6):355-65
pubmed: 21614641
Lung Cancer. 1996 Nov;15(3):311-23
pubmed: 8959677
Nat Protoc. 2008;3(6):1101-8
pubmed: 18546601
Biochem Biophys Res Commun. 2002 Jan 18;290(2):763-9
pubmed: 11785965
FASEB J. 1993 Dec;7(15):1475-82
pubmed: 8262332
Stem Cells Int. 2018 Jun 6;2018:5340756
pubmed: 29977307
Am J Vet Res. 2007 Oct;68(10):1095-105
pubmed: 17916017
J Am Vet Med Assoc. 2005 May 1;226(9):1529-37
pubmed: 15882006
Stem Cell Res Ther. 2018 Jul 4;9(1):178
pubmed: 29973295
Matrix Biol. 2000 Dec;19(7):615-22
pubmed: 11102750
J Craniomaxillofac Surg. 2015 Dec;43(10):2144-51
pubmed: 26541747
J Biol Chem. 1946 Jul;164:321-9
pubmed: 20989492
Cell Stem Cell. 2008 Apr 10;2(4):313-9
pubmed: 18397751
Metabolism. 2004 May;53(5):632-7
pubmed: 15131769
Pathobiology. 2016;83(5):258-66
pubmed: 27225269
Surg Pathol Clin. 2019 Mar;12(1):21-33
pubmed: 30709444
ANZ J Surg. 2009 Apr;79(4):235-44
pubmed: 19432707
Br J Dermatol. 2009 Oct;161(4):819-25
pubmed: 19558598
Am J Stem Cells. 2015 Mar 15;4(1):1-12
pubmed: 25973326
J Orthop Res. 1997 Jul;15(4):546-57
pubmed: 9379264
J Bone Miner Res. 2002 Aug;17(8):1486-97
pubmed: 12162503
Stem Cells Dev. 2010 Oct;19(10):1449-70
pubmed: 20486777
Virchows Arch. 1995;427(4):353-63
pubmed: 8548119
Nat Protoc. 2006;1(3):1112-6
pubmed: 17406391
J Vis Exp. 2017 Feb 24;(120):
pubmed: 28287507
Aesthetic Plast Surg. 2007 Sep-Oct;31(5):574-8
pubmed: 17576503
Oral Surg Oral Med Oral Pathol Oral Radiol. 2016 Jul;122(1):e8-e13
pubmed: 26652892
Diabetes. 2012 Jul;61(7):1691-9
pubmed: 22596050
J Cutan Pathol. 2007 Oct;34(10):788-92
pubmed: 17880585
J Cell Physiol. 2001 Oct;189(1):54-63
pubmed: 11573204
Stem Cells Transl Med. 2014 Feb;3(2):206-17
pubmed: 24361924
J Korean Neurosurg Soc. 2013 Dec;54(6):518-20
pubmed: 24527197