Optimizing Osteogenic Differentiation of Ovine Adipose-Derived Stem Cells by Osteogenic Induction Medium and FGFb, BMP2, or NELL1 In Vitro

Abstract

Although adipose-derived stromal cells (ADSCs) have been a major focus as an alternative to autologous bone graft in orthopedic surgery, bone formation potential of ADSCs is not well known and cytokines as osteogenic inducers on ADSCs are being investigated. This study aimed at isolating ADSCs from ovine adipose tissue (AT) and optimizing osteogenic differentiation of ovine ADSCs (oADSC) by culture medium and growth factors. Four AT samples were harvested from two female ovine (Texel/Gotland breed), and oADSCs were isolated and analyzed by flow cytometry for surface markers CD29, CD44, CD31, and CD45. Osteogenic differentiation was made in vitro by seeding oADSCs in osteogenic induction medium (OIM) containing fibroblast growth factor basic (FGFb), bone morphogenetic protein 2 (BMP2), or NEL-like molecule 1 (NELL1) in 4 different dosages (1, 10, 50, and 100 ng/ml, respectively). Basic medium (DMEM) was used as control. Analysis was made after 14 days by Alizarin red staining (ARS) and quantification. This study successfully harvested AT from ovine and verified isolated cells for minimal criteria for adipose stromal cells which suggests a feasible method for isolation of oADSCs. OIM showed significantly higher ARS to basic medium, and FGFb 10 ng/ml revealed significantly higher ARS to OIM alone after 14 days.

AML‑derived mesenchymal stem cells upregulate CTGF expression through the BMP pathway and induce K562‑ADM fusiform transformation and chemoresistance

“…The primary AML-MSCs were then obtained. The cultured AML-MSCs were cultured in osteogenic induction medium (500 µl vitamin C, 1 ml β-glycerophosphate and 10 µl dexamethasone in 100 ml DMEM) (45), followed by alkaline phosphatase assay and Alizarin Red staining to verify its ability to differentiate into osteoblasts.…”

Adipose-Derived Stem Cells Inhibited the Proliferation of Bladder Tumor Cells by S Phase Arrest and Wnt/β-Catenin Pathway

“…Adipose-derived stem cells (ADSCs) isolated from the stromal vascular fraction of adipose tissue share the same characteristics as mesenchymal stem cells (MSCs) and can differentiate into adipogenic, myogenic, osteogenic, chondrogenic, and neurogenic cells (Nielsen et al, 2018; Zachar et al, 2011). Currently, many studies have linked many important growth factors, cytokines, and chemokines secreted by ADSCs and MSCs to cancer development and progression (Chen et al, 2018; Chu et al, 2018; Gazdic et al, 2017; Lu et al, 2016).…”

Extracellular vesicles derived from human dental pulp stem cells promote osteogenesis of adipose-derived stem cells via the MAPK pathway

“…HADSCs have been shown to be a promising and appealing source of MSCs for BTE due to their abundance, the fact that they are less invasive to harvest, and their efficient growth in culture. 41 Accumulating research have proven that hADSCs are highly sensitive to the culture environment, which may affect their osteogenic differentiation. 42 Previous in vitro studies verified the superior osteogenesis of BMSCs compared to ADSCs 43 ; however, controversy remains over which cells exhibit better osteogenic differentiation potential in vivo.…”