Acta chirurgiae orthopaedicae
et traumatologiae Čechoslovaca

Acta Chir Orthop Traumatol Cech. 2011; 78(2):138-144 | DOI: 10.55095/achot2011/022

Výsledky srovnání chondrogenní diferenciace mezenchymálních kmenových buněk získaných z tukové tkáně s kultivovanými chondrocyty a mezenchymálními kmenovými buňkami z kostní dřeněPůvodní práce

V. HAVLAS^1,*, P. KOS¹, P. JENDELOVÁ^2,3, P. LESNݲ, T. TRČ¹, E. SYKOVÁ^2,3

¹ Ortopedická klinika UK 2. LF a FN Motol, dětská a dospělá ortopedie a traumatologie, Praha

² Centrum buněčné terapie a tkáňových náhrad 2. LF UK a FN Motol, Praha

³ Ústav experimentální medicíny AV ČR, Praha

PURPOSE OF THE STUDY:
Many congenital and acquired disorders as well as sequelae of injury are associated with articular cartilage degeneration, which adversely affects the patient's quality of life. The currently used cell therapy with cultured chondrocytes has its disadvantages due to a process of de-differentiation of chondrocytes during cultivation. We believe that the mesenchymal stem cell therapy offers a new treatment options.

MATERIAL AND METHODS:
The adult mesenchymal stem cells (MSCs) for chondrocyte differentiation are usually obtained from bone marrow mesenchymal stem cells (BMSCs). In this study these cells were compared with mesenchymal stem cells derived from adipose tissue (AMSCs). The aim of the study was to verify the ability of human BMSCs and AMSCs to differentiate into chondrocytes in vitro in the presence or absence of transforming growth factor beta (TGF-β1). Human BMSCs and AMSCs were collected from healthy donors during orthopaedic surgeries, in vitro cultured in three passages to obtain the required quantity of cells. A pellet culture system was used for chondrocyte differentiation.

RESULTS:
At 21 days of cultivation, cell aggregates grown in the chondrogenic medium were larger than those cultured in the control medium. Both the BMSCs and AMSCs pellet cultures showed spontaneous chondrogenesis. Histological staining with haematoxylin and eosin and Masson's trichrome stains, as well as immunohistochemical staining to detect type II collagen revealed no apparent differences between the pellet cultures with TGF-β1 presence and those without it. The real-time RT-PCR detected expression of the type II collagen gene in all tested cultures. In the BMSCs pellet culture only, TGF-β1 presence resulted in a decrease in type I collagen mRNA levels and in an increase in type II collagen mRNA values.

DISCUSSION:
Our results showed an in vitro chondrogenic potential of mature human mesenchymal stem cells derived from both bone marrow and adipose tissue. In agreement with the relevant literature data, we suggest that both cell types have an equal prospect for use in cartilage tissue engineering.

Klíčová slova: bone marrow, adipose tissue, transforming growth factor beta, cartilage, pellet culture system

Zveřejněno: 1. duben 2011  Zobrazit citaci