PMSC pellets were obtained by centrifuging 2.5 × 104 cells at 250 × g for 5 min in 15 ml polypropylene conical tubes as reported12 (link). Pellets were cultured for 14 days in 0.5 ml of serum-free MesenCult™-Chondrogenic differentiation medium (Stem cell Technologies, Vancouver, Canada) containing soluble JAG1 (10 µg/ml) or lgG (10 µg/ml) peptides. Harvested pellets were fixed in 4% paraformaldehyde (PFA), dehydrated, and embedded in paraffin. Serial sections of 5 µm were stained with alcian blue/orange G. Immunohistochemical (IHC) analysis for Col-II was performed as described previously12 (link).
Mesencult chondrogenic differentiation medium
Manufactured by STEMCELL
Sourced in Canada
MesenCult™-Chondrogenic differentiation medium is a defined, serum-free culture medium designed to support the chondrogenic differentiation of mesenchymal stem/stromal cells (MSCs) in vitro.
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Lab products found in correlation
2 protocols using mesencult chondrogenic differentiation medium
1
Chondrogenic Differentiation of Mesenchymal Stem Cells
2
Perivascular Niche Self-Renewal Evaluation
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Monolayer cell chondrogenesis was assessed by culture in MesenCult Chondrogenic Differentiation medium (StemCell Technologies Inc, Vancouver, British Colombia). Medium was changed every 3 days.
Cells were washed with PBS, and fixed with 4% PFA for 15 minutes.
Cultures were then stained with alcian blue at 21 days of differentiation. Three wells per group were used. To document potential self-renewal of the perivascular niche in vivo, 5 × 10 5 PDGFRα + or PDGFRα -cells (passage 5) were percutaneously injected within the mid-substance of the inguinal fat pad using a 8-μL PBS carrier. Three NOD/SCID mice per group were used, and animals were sacrificed 2 weeks after transplantation. Inguinal fat pads were sectioned at 20 μm thickness and stained with CD31 and CD34 antibodies as per the above descriptions.
Cells were washed with PBS, and fixed with 4% PFA for 15 minutes.
Cultures were then stained with alcian blue at 21 days of differentiation. Three wells per group were used. To document potential self-renewal of the perivascular niche in vivo, 5 × 10 5 PDGFRα + or PDGFRα -cells (passage 5) were percutaneously injected within the mid-substance of the inguinal fat pad using a 8-μL PBS carrier. Three NOD/SCID mice per group were used, and animals were sacrificed 2 weeks after transplantation. Inguinal fat pads were sectioned at 20 μm thickness and stained with CD31 and CD34 antibodies as per the above descriptions.
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