Protocols for in vitro Differentiation of Human Mesenchymal Stem Cells into Osteogenic, Chondrogenic and Adipogenic Lineages

Abstract

Mesenchymal stem cells (MSC) possess high plasticity and the potential to differentiate into several different cell types; this characteristic has implications for cell therapy and reparative biotechnologies. MSC have been originally isolated from the bone marrow (BM-MSC), but they have been found also in other tissues such as adipose tissue, cord blood, synovium, skeletal muscle, and lung. MSC are able to differentiate in vitro and in vivo into several cell types such as bone, osteocytes, chondrocytes, adipocytes, and skeletal myocytes, just to name a few.During the last two decades, an increasing number of studies have proven the therapeutic potential of MSC for the treatment of neurodegenerative diseases, spinal cord and brain injuries, cardiovascular diseases, diabetes mellitus, and diseases of the skeleton. Their immuno-privileged profile allows both autologous and allogeneic use. For all these reasons, the scientific appeal of MSC is constantly on the rise.The identity of MSC is currently based on three main criteria: plastic-adherence capacity, defined epitope profile, and capacity to differentiate in vitro into osteocytes, chondrocytes, and adipocytes. Here, we describe standard protocols for the differentiation of BM-MSC into the osteogenic, chondrogenic, and adipogenic lineages.

Mesenchymal Stromal/Stem Cells in Regenerative Medicine and Tissue Engineering

“…Differentiation protocols for driving differentiation toward these lineages have been routinely utilized and extensively optimized [52, 53]. For example, osteogenesis typically involves the use of dexamethasone, β -glycerolphosphate, and ascorbic acid.…”

Gestational diabetes impacts fetal precursor cell responses with potential consequences for offspring

“…To determine the multilineage differentiation capacity of AMSCs, specific differentiation conditions were used to trigger cell differentiation into adipocytes, chondrocytes, and osteocytes as described . Differentiated cells were stained with Oil Red O, Alcian Blue or Alizarin Red, respectively, and observed in a bright‐field microscope (ZEISS Axio Vert A1, Oberkochen, Germany).…”

“…To determine the multilineage differentiation capacity of AMSCs, specific differentiation conditions were used to trigger cell differentiation into adipocytes, chondrocytes, and osteocytes as described. 17…”

Effect of Photobiomodulation Therapy on the Increase of Viability and Proliferation of Human Mesenchymal Stem Cells

“…After 24 hours, the medium was changed to an adipogenesis differentiation medium and the cells cultured as above for 7–10 days before being examined by confocal microscopy—see “Confocal microscopy imaging” section below. The protocol again was that of Ciuffreda et al …”

“…To differentiate MSCs into chondrogenic, osteogenic, and adipogenic lineages, StemPro Differentiation Kits (Gibco) were used. After reaching 60-80% confluence, low-passage cells (passages 3-4) were detached using TrypLE™ Express (Gibco), washed with PBS, and resuspended in CCM by using the protocol of Ciuffreda et al [39] Adipogenesis differentiation. MSCs suspended in CCM were transferred to a 96-well plate (black with glass bottom; Sensoplate) at 1 × 10 4 cells/cm 2 .…”