open access

Vol 3, No 3 (2018)
Original article
Published online: 2018-11-02
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Does the conditioned medium trigger the adipose-derived mesenchymal stem cells differentiation into Chondrocytes?

Łukasz Kaźmierski1, Anna Bajek1, Robert Dębski2, Magdalena Bodnar3, Krzysztof Roszkowski4
·
Medical Research Journal 2018;3(3):148-152.
Affiliations
  1. Department of Tissue Engineering, Collegium Medicum Bydgoszcz, Nicolaus Copernicus University, Karłowicza 24, 85-092 Bydgoszcz, Poland
  2. Department of Pediatric Hematology and Oncology, Collegium Medicum Bydgoszcz, Nicolaus Copernicus University, Curie Skłodowskiej 9, 85-094 Bydgoszcz, Poland
  3. Department of Clinical Pathomorphology, Collegium Medicum Bydgoszcz, Nicolaus Copernicus University, Curie Skłodowskiej 9, 85-094 Bydgoszcz, Poland
  4. Department of Oncology, Radiotherapy and Ginecologic Oncology, Collegium Medicum Bydgoszcz, Nicolaus Copernicus University,, Romanowskiej 2, 85-796 Bydgoszcz, Poland

open access

Vol 3, No 3 (2018)
ORIGINAL ARTICLES
Published online: 2018-11-02

Abstract

Background: Cartilage is an avascular tissue that has limited regenerative capacity. Stem cells, especially
mesenchymal stem cells (MSCs), have been regarded as a promising cell source for cartilage repair due
to their multi-lineage differentiation potential. Induction of mesenchymal stem cells into functional chondrocytes
may resolve the above problems in the cartilage repair. However, the imperative is to develop
effective strategies for chondrogenic differentiation. The aim of this study is to evaluate if the induction of
adipose-derived mesenchymal stem cells (ADSCs) differentiation toward chondrocytes is possible with
the use of conditioned medium derived from chondrocytes culture.


Material and methods: Conditioned medium (CM) from Normal chondrocyte cell line CRL 2648 was used to culture ADSCs, at the same time ADSCs with standard growth medium were used as a control. During this experiment, the phenotype was assessed using flow cytometry (CD90, CD44, CD45 and CD45), and Immunohistochemical
staining was used to determine differentiation (anti-aggrecan, anti-CD15, anti-collagen II, anti-TRA-2-49).

Results: ADSCs propagated rapidly in vitro and formed a homogenous fibroblast-like morphology. 0% of tested
cells showed the expression of CD90 and CD44, while more than 90% did not reveal the expression of
CD34 and CD45. Results obtained in this study showed weak cytoplasmatic expression of aggrecan
and alkaline phosphatase (TRA protein) and lack of CD151 expression. However, the majority of ADSCs
expanded with CM expressed type II collagen indicating that the chondrocyte-secreted factors induced
chondrogenic commitment during expansion.


Conclusions: In the natural environment of living organisms, the chondrogenic differentiation of stem cells involve multiple signalling pathways. We can, to an extent mimic these signals in vitro however, for the clinical
purposes it is very important to develop well-defined and efficient in vitro protocols. Our results indicate
that CM may regulate and induce differentiation of ADSCs into the chondrocyte lineage and can serve
also as an in vitro model for studying specific lineage commitment.

Abstract

Background: Cartilage is an avascular tissue that has limited regenerative capacity. Stem cells, especially
mesenchymal stem cells (MSCs), have been regarded as a promising cell source for cartilage repair due
to their multi-lineage differentiation potential. Induction of mesenchymal stem cells into functional chondrocytes
may resolve the above problems in the cartilage repair. However, the imperative is to develop
effective strategies for chondrogenic differentiation. The aim of this study is to evaluate if the induction of
adipose-derived mesenchymal stem cells (ADSCs) differentiation toward chondrocytes is possible with
the use of conditioned medium derived from chondrocytes culture.


Material and methods: Conditioned medium (CM) from Normal chondrocyte cell line CRL 2648 was used to culture ADSCs, at the same time ADSCs with standard growth medium were used as a control. During this experiment, the phenotype was assessed using flow cytometry (CD90, CD44, CD45 and CD45), and Immunohistochemical
staining was used to determine differentiation (anti-aggrecan, anti-CD15, anti-collagen II, anti-TRA-2-49).

Results: ADSCs propagated rapidly in vitro and formed a homogenous fibroblast-like morphology. 0% of tested
cells showed the expression of CD90 and CD44, while more than 90% did not reveal the expression of
CD34 and CD45. Results obtained in this study showed weak cytoplasmatic expression of aggrecan
and alkaline phosphatase (TRA protein) and lack of CD151 expression. However, the majority of ADSCs
expanded with CM expressed type II collagen indicating that the chondrocyte-secreted factors induced
chondrogenic commitment during expansion.


Conclusions: In the natural environment of living organisms, the chondrogenic differentiation of stem cells involve multiple signalling pathways. We can, to an extent mimic these signals in vitro however, for the clinical
purposes it is very important to develop well-defined and efficient in vitro protocols. Our results indicate
that CM may regulate and induce differentiation of ADSCs into the chondrocyte lineage and can serve
also as an in vitro model for studying specific lineage commitment.

Get Citation

Keywords

cartilage injury, conditioned medium, mesenchymal stem cells, chondrocytes, adipose-derived mesenchymal stem cells

About this article
Title

Does the conditioned medium trigger the adipose-derived mesenchymal stem cells differentiation into Chondrocytes?

Journal

Medical Research Journal

Issue

Vol 3, No 3 (2018)

Article type

Original article

Pages

148-152

Published online

2018-11-02

Page views

980

Article views/downloads

790

DOI

10.5603/MRJ.a2018.0025

Bibliographic record

Medical Research Journal 2018;3(3):148-152.

Keywords

cartilage injury
conditioned medium
mesenchymal stem cells
chondrocytes
adipose-derived mesenchymal stem cells

Authors

Łukasz Kaźmierski
Anna Bajek
Robert Dębski
Magdalena Bodnar
Krzysztof Roszkowski

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