The effective mode of growth and differentiation factor-5 in promoting the chondrogenic differentiation of adipose-derived stromal cells

Abstract

Our study aimed to find out the most effective mode for chondrogenic differentiation based on time, dose and culture method. ADSCs were cultured and identified by CD44, CD49d, and CD106 immumohistochemical staining method, and their differentiation potential to chondrocyte were detected by Alizarin red staining. ADSCs induced by different concentrations of GDF-5 for chondrogenic differentiation were detected by blue and toluidine blue staining and collagen type II and X immumohistochemical staining. The expression of collagen I, II, X and aggrecan gene in GDF-induced ADSCs cultured in 2- and 3-dimension was identified by real-time PCR. Cell microstructure and proliferation in three-dimensional scaffolds at day 7, 14, 21 and 28 were analyzed by scanning electron microscopy and MTS assay. The ADSCs were successfully identified by CD44 and CD49d, and their differentiation potential was detected by Alizarin red staining. Real-time PCR showed that collagen and aggrecan were expressed at high levels in 100 or 200 ng/mL GDF-5 treated cells. The collagen types (I, II) and aggrecan genes were higher expressed in GDF-5 induced scaffold group than that in monolayer group. MTS showed that the cell counts were not significantly different among different treated time. Both collagen type II and aggrecan gene were highly expressed at day 14, while collagen types I and X gene expressions peaked at day 21 and 28. The 100 ng/mL GDF-5 is effective and cost-effective for chondrogenic differentiation when cultured at day 14 in vitro under three-dimensional culture conditions.

Therapeutic potential of growth differentiation factors in the treatment of degenerative disc diseases

“…Both GDF5 and GDF6 have also been shown to induce the expression of general chondrogenic genes (Sex determining region Y‐Box9 (SOX9); type II collagen; ACAN) and, importantly, to also induce NP‐specific genes (KRT18, KRT19, CA12, Brachyury (T), CD24, HIF1α, Glut‐1 and SHH) in MSCs and adipose‐derived stem cells (ASCs) (Table ). When GDF5 is added as a media supplement to high‐density cultures of MSCs or ASCs, chondrogenic differentiation is increased; however, several reports also note a significant increase in markers of hypertrophy and ossification including alkaline phosphatase, collagen types I and X, and osteopontin . These characteristics indicate a progression in expression reflective of chondrogenic hypertrophy toward endochondral ossification, which is undesirable for NP populations.…”

Fabrication of a Cu/Zn co-incorporated calcium phosphate scaffold-derived GDF-5 sustained release system with enhanced angiogenesis and osteogenesis properties

“…Previous studies have found that GDF-5 promoted the recruitment of mesenchymal stem cells (MSCs) and their differentiation into chondrocytes, thus accelerating chondrogenesis and joint formation. 15,16 In recent years, GDF-5 has shown great application potential in bone tissue repair. GDF-5 deciency resulted in delayed bone healing, 17 whereas its addition promoted MSCs osteogenic differentiation and VEGF expression.…”

Ginkgo biloba Extract 50 (GBE50) Ameliorates Insulin Resistance, Hepatic Steatosis and Liver Injury in High Fat Diet-Fed Mice

“…The procedure of immunohistochemical staining was described as previously. 14 Briefly, paraffin-embedded liver sections (5 μm thickness) were deparaffinized and rehydrated, then disposed with 3% H 2 O 2 for 10 min followed by blocking with 5% donkey serum for 30 min at room temperature, finally, incubated with primary antibodies against F4/80 (macrophage marker; cat No. ab16911; 1:200, Abcam, Cambridge, MA, USA), CD45 (leukocyte marker; cat No.…”