AB0094 DIFFERENTIATION OF ADIPOSE DERIVED MESENCHYMAL STEM CELLS OBTAINED FROM PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS ANKYLOSING SPONDYLITIS AND SYSTEMIC SCLEROSIS

Background:

Cartilage and bone destruction occurs in many rheumatic diseases. While the use of biologic drugs may delay the destruction but still it cannot be averted. Adipose tissue is an easy accessible and rich source of MSCs. Application of mesenchymal stem/stromal cells (MSCs) may be promising option for successful tissue regeneration in therapy of ankylosing spondylitis (AS) systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) patients.

Objectives:

The aim of this study was to compare differentiation potential of ASCs obtained from AS, SLE, SSc patients and ASCs line originating from healthy volunteers (hASCs).The phenotype of these cells has been also analysed.

Methods:

ASCs obtained from AS (n=9), SLE (n=10), SSc (n=10) patients and 5 commercially available hASC lines were used in study. Cells in passage 4 were used in each experiment. Phenotype of ASCs was evaluated by flow cytometry. Differentiation was made by using osteogenic, chondrogenic or adipogenic media. At the end of differentiation process cells were harvested and total RNA was isolated. Relative quantification (RQ) of gene expression level were calculated by the 2−ΔΔct method. After 4 weeks of chondrogenic differentiation, mRNA level of SOX9 and aggrecan (ACAN) mRNA has been evaluated by RT-PCR. Additionally glycosaminoglycan (GAG) deposition was analysed by alcian blue staining. After 2 weeks of osteogenic differentiation, expression of RUNX-2, collagen 1a1 (COL1a1) and osteopontin (OPN) mRNA has been determined. Calcium deposition has been determined by alizarin red staining. After 3 weeks of adipogenic differentiation expression of CCAAT/enhancer-binding protein (C/EBP), peroxisome proliferator activated receptor-γ (PPAR-γ) and fatty acid binding protein 4 (FABP4) was determined.

Results:

All ASCs cultured in osteogenic medium showed calcium deposition. The expression of RUNX-2 and OPN mRNA was significantly higher in AS-ASCs. Cells obtained from SLE and SSc revealed significantly lower expression of COL1a1 than hASCs lines. The results of alcian blue staining showed chondrogenesis of cells obtained from all patients types. No statistically significant differences between AS, SLE, SSc and hASCa lines were observed in SOX9 mRNA expression. However, all patients derived cells expressed a lower of ACAN mRNA level. Deposition of oil droplets in cytoplama was observed in all cells cultured in adipogenic medium. There were no differences in expression of C/EBP and FAB4. Cell derived from SLE and AS expressed significantly higher level of PPARγ.

Conclusion:

AS, SLE and SSc ASCs have phenotype comparable with hASCs lines. The patients derived ASCs are mighty to differentiate into any of the 3 cell types, although the process is altered.

Acknowledgement:

This work was sponsored by grant No 2016/21/B/NZ5/00500 from National Science Centre, Poland

Disclosure of Interests:

Ewa Kuca-Warnawin: None declared, Magdalena Plebanczyk: None declared, Anna Wajda: None declared, Krzysztof Bonek: None declared, Piotr Głuszko: None declared, Piotr Szczęsny: None declared, Marzena Olesińska Consultant for: F. Hoffmann-La Roche, Wlodzimierz Maśliński: None declared, Ewa Kontny: None declared