Activin/BMP2 chimeric ligands direct adipose-derived stem cells to chondrogenic differentiation

Macarena Peran; Sergio Ruiz; Witek Kwiatkowski; Juan Antonio Marchal; Sheng Lian Yang; Antonia Aranega; Senyon Choe; Juan Carlos Izpisua Belmonte

doi:10.1016/j.scr.2013.02.002

Activin/BMP2 chimeric ligands direct adipose-derived stem cells to chondrogenic differentiation

Macarena Peran, Sergio Ruiz, Witek Kwiatkowski, Juan Antonio Marchal, Sheng Lian Yang, Antonia Aranega, Senyon Choe^*, Juan Carlos Izpisua Belmonte

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

Human adipose derived stem cells (hASCs) can be easily isolated and their plasticity has been well characterized. Several TGF-β superfamily ligands can direct hASCs towards chondrocytes. However, these ligands are difficult to purify and expensive. We have developed a library of Activin/BMP2 chimeric ligands (AB2 ligands) by systematically mixing their sequence segments and have tested their chondrogenic potential in hASCs. Cells cultured in monolayer or in a pellet culture system were incubated with a chemically defined medium supplemented with the chimeric ligands for 4 or 6. weeks and showed higher expression levels of type II collagen, aggrecan, and Sox9 mRNAs when compared with control and non-treated cells. Moreover, toluidine blue, alcian blue, and Masson's trichrome staining was markedly increased in treated cells, both in cell pellet and monolayer assays. In addition, immunohistochemical staining for detection of type I collagen, type II collagen, and Sox 9 demonstrated the acquisition of a chondrogenic phenotype in both culture systems. We present here an inexpensive and robust protocol for differentiation of hASCs towards chondrocytes in a reproducible and highly efficient manner. The AB2 ligands employed are easily produced and have properties that may become useful in cell therapy.

Original language	English (US)
Pages (from-to)	464-476
Number of pages	13
Journal	Stem Cell Research
Volume	10
Issue number	3
DOIs	https://doi.org/10.1016/j.scr.2013.02.002
State	Published - May 2013
Externally published	Yes

Bibliographical note

Funding Information:
Macarena Peran was supported first by the Spanish Ministry of Education through the National Human Resources Mobility National Plan I-D + i 2008–2011 and continued with a grant from Jaen University, Spain . J.A.M.: Junta de Andalucía, excellence project number CTS-6568. Work in the laboratory of J.C.I.B. was supported by grants from TERCEL-ISCIII-MINECO, Fundacion Cellex, CIBER-BBN , The Leona M. and Harry B. Helmsley Charitable Trust, G. Harold and Leila Y. Mathers Charitable Foundation and The Ellison Medical Foundation.

ASJC Scopus subject areas

Developmental Biology
Cell Biology

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10.1016/j.scr.2013.02.002

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@article{167649cf73094bdbb749a0d6321a71cb,

title = "Activin/BMP2 chimeric ligands direct adipose-derived stem cells to chondrogenic differentiation",

abstract = "Human adipose derived stem cells (hASCs) can be easily isolated and their plasticity has been well characterized. Several TGF-β superfamily ligands can direct hASCs towards chondrocytes. However, these ligands are difficult to purify and expensive. We have developed a library of Activin/BMP2 chimeric ligands (AB2 ligands) by systematically mixing their sequence segments and have tested their chondrogenic potential in hASCs. Cells cultured in monolayer or in a pellet culture system were incubated with a chemically defined medium supplemented with the chimeric ligands for 4 or 6. weeks and showed higher expression levels of type II collagen, aggrecan, and Sox9 mRNAs when compared with control and non-treated cells. Moreover, toluidine blue, alcian blue, and Masson's trichrome staining was markedly increased in treated cells, both in cell pellet and monolayer assays. In addition, immunohistochemical staining for detection of type I collagen, type II collagen, and Sox 9 demonstrated the acquisition of a chondrogenic phenotype in both culture systems. We present here an inexpensive and robust protocol for differentiation of hASCs towards chondrocytes in a reproducible and highly efficient manner. The AB2 ligands employed are easily produced and have properties that may become useful in cell therapy.",

author = "Macarena Peran and Sergio Ruiz and Witek Kwiatkowski and Marchal, {Juan Antonio} and Yang, {Sheng Lian} and Antonia Aranega and Senyon Choe and {Izpisua Belmonte}, {Juan Carlos}",

note = "Funding Information: Macarena Peran was supported first by the Spanish Ministry of Education through the National Human Resources Mobility National Plan I-D + i 2008–2011 and continued with a grant from Jaen University, Spain . J.A.M.: Junta de Andaluc{\'i}a, excellence project number CTS-6568. Work in the laboratory of J.C.I.B. was supported by grants from TERCEL-ISCIII-MINECO, Fundacion Cellex, CIBER-BBN , The Leona M. and Harry B. Helmsley Charitable Trust, G. Harold and Leila Y. Mathers Charitable Foundation and The Ellison Medical Foundation.",

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AU - Peran, Macarena

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AU - Yang, Sheng Lian

AU - Aranega, Antonia

AU - Choe, Senyon

AU - Izpisua Belmonte, Juan Carlos

N1 - Funding Information: Macarena Peran was supported first by the Spanish Ministry of Education through the National Human Resources Mobility National Plan I-D + i 2008–2011 and continued with a grant from Jaen University, Spain . J.A.M.: Junta de Andalucía, excellence project number CTS-6568. Work in the laboratory of J.C.I.B. was supported by grants from TERCEL-ISCIII-MINECO, Fundacion Cellex, CIBER-BBN , The Leona M. and Harry B. Helmsley Charitable Trust, G. Harold and Leila Y. Mathers Charitable Foundation and The Ellison Medical Foundation.

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N2 - Human adipose derived stem cells (hASCs) can be easily isolated and their plasticity has been well characterized. Several TGF-β superfamily ligands can direct hASCs towards chondrocytes. However, these ligands are difficult to purify and expensive. We have developed a library of Activin/BMP2 chimeric ligands (AB2 ligands) by systematically mixing their sequence segments and have tested their chondrogenic potential in hASCs. Cells cultured in monolayer or in a pellet culture system were incubated with a chemically defined medium supplemented with the chimeric ligands for 4 or 6. weeks and showed higher expression levels of type II collagen, aggrecan, and Sox9 mRNAs when compared with control and non-treated cells. Moreover, toluidine blue, alcian blue, and Masson's trichrome staining was markedly increased in treated cells, both in cell pellet and monolayer assays. In addition, immunohistochemical staining for detection of type I collagen, type II collagen, and Sox 9 demonstrated the acquisition of a chondrogenic phenotype in both culture systems. We present here an inexpensive and robust protocol for differentiation of hASCs towards chondrocytes in a reproducible and highly efficient manner. The AB2 ligands employed are easily produced and have properties that may become useful in cell therapy.

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Activin/BMP2 chimeric ligands direct adipose-derived stem cells to chondrogenic differentiation

Abstract

Bibliographical note

ASJC Scopus subject areas

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