The effects of bone morphogenetic protein 6, dexamethasone and human serum on chondrogenic differentiation of human adipose stem cells in vitro
PÄÄKKÖNEN, RAMI (2013)
2013
Biokemia - Biochemistry
Biolääketieteellisen teknologian yksikkö - Institute of Biomedical Technology
This publication is copyrighted. You may download, display and print it for Your own personal use. Commercial use is prohibited.
Hyväksymispäivämäärä
2013-01-14
Julkaisun pysyvä osoite on
https://urn.fi/urn:nbn:fi:uta-1-23543
https://urn.fi/urn:nbn:fi:uta-1-23543
Tiivistelmä
Background and aims: Damaged articular cartilage is a common cause of pain, lowered mobility, lowered working capability and reduced quality of life. Injured articular cartilage hardly heals and the current treatments have only proven applicable for a fraction of patients, hence the great demand for a novel treatment. Mesenchymal stem cells from human adipose tissue are self-proliferative and they can undergo multilineage differentiation. In this study, we aim to examine the chondrogenic culture conditions and media compositions for adipose stem cells and to seek possible improvements.
Methods: Human adipose stem cells were chondrogenically differentiated with six media containing various combinations and doses of putative chondrogenic bioactive factors, namely TGF-β1, BMP-6, dexamethasone and human serum. After 2 and 4 weeks in three-dimensional high cell-density aggregate culture the expression of osteogenesis- and chondrogenesis related genes was analyzed with commercial Q-RT-PCR array plates. Histological stainings and immunohistochemistry were used to verify the accumulation of cartilaginous extracellular matrix.
Results: None of the media supplements proved to induce chondrogenesis exclusively. BMP-6 resulted in mixed chondrogenic and osteogenic differentiation, although a high dose of BMP-6 did suppress the hypertrophic markerCOL10A1. Dexamethasone boosted the expression of a variety of ECM related genes, namelyCOL1A2,COL3A1,COL11A1,COL12A1,COMP,SERPINH1andALP. When combined, a high dose of BMP-6 and dexamethasone resulted in putative adipogenic differentiation. Human serum proved unnecessary, or even detrimental for chondrogenesis.
Conclusions: We conclude that the growth factor combinations assayed here proved suboptimal for chondrogenesis; the cells underwent concomitant osteogenesis and even putative adipogenesis was observed. However, with further refinement of the chondrogenic culturing conditions the differentiation of human adipose stem cells should be even more controllable, and accordingly cartilage tissue engineering with adipose stem cells would seem feasible.
Keywords: Tissue engineering, Mesenchymal stem cells, Chondrogenesis, TGF-beta superfamily proteins, Bone morphogenetic protein 6, Hypertrophy Kudosteknologia, Mesenkymaaliset kantasolut, Rustoerilaistus, TGF-beta superperheen proteiinit, Luun morfogeneettinen proteiini 6, Hypertrofia
Methods: Human adipose stem cells were chondrogenically differentiated with six media containing various combinations and doses of putative chondrogenic bioactive factors, namely TGF-β1, BMP-6, dexamethasone and human serum. After 2 and 4 weeks in three-dimensional high cell-density aggregate culture the expression of osteogenesis- and chondrogenesis related genes was analyzed with commercial Q-RT-PCR array plates. Histological stainings and immunohistochemistry were used to verify the accumulation of cartilaginous extracellular matrix.
Results: None of the media supplements proved to induce chondrogenesis exclusively. BMP-6 resulted in mixed chondrogenic and osteogenic differentiation, although a high dose of BMP-6 did suppress the hypertrophic markerCOL10A1. Dexamethasone boosted the expression of a variety of ECM related genes, namelyCOL1A2,COL3A1,COL11A1,COL12A1,COMP,SERPINH1andALP. When combined, a high dose of BMP-6 and dexamethasone resulted in putative adipogenic differentiation. Human serum proved unnecessary, or even detrimental for chondrogenesis.
Conclusions: We conclude that the growth factor combinations assayed here proved suboptimal for chondrogenesis; the cells underwent concomitant osteogenesis and even putative adipogenesis was observed. However, with further refinement of the chondrogenic culturing conditions the differentiation of human adipose stem cells should be even more controllable, and accordingly cartilage tissue engineering with adipose stem cells would seem feasible.
Keywords: Tissue engineering, Mesenchymal stem cells, Chondrogenesis, TGF-beta superfamily proteins, Bone morphogenetic protein 6, Hypertrophy Kudosteknologia, Mesenkymaaliset kantasolut, Rustoerilaistus, TGF-beta superperheen proteiinit, Luun morfogeneettinen proteiini 6, Hypertrofia