**3.2 3-D cell pellet culture and chondrogenesis of human BM-MSCs**

Chondrogenic medium: 95% DMEM high-glucose medium (Sigma-Aldrich, St Louis, MO), 1% 1 × ITS+1 solution (BD Biosciences, San Jose, CA), 1% Pen-Strep,

#### **Figure 2.**

*Chondrogenic culture of human BM-MSCs. Representative images (10×) of immunohistochemical staining of collage II (A) and aggrecan (B) in pellet culture samples at day 17 during chondrogenesis [29].*

100 μg/mL sodium pyruvate (Invitrogen, Carlsbad, CA), 50 μg/mL AsAP, 40 μg/ mL L-proline (Sigma-Aldrich, St Louis, MO), 0.1 μM dexamethasone, and 10 ng/mL recombinant human TGF-β3 (Lonza, Walkersville, MD).

Passage 4 human BM-MSCs are used for chondrogenic differentiation. Chondrogenic differentiation is induced by chondrogenic medium (**Figure 2**).


Day 1: Fixation and dehydration


*Isolation and Expansion of Mesenchymal Stem/Stromal Cells, Functional Assays and Long-Term… DOI: http://dx.doi.org/10.5772/intechopen.100286*


#### **3.3** *In vitro* **adipogenesis of human MSCs**

Adipogenic medium: DMEM (1 g/L glucose), 10% FBS, 1% penicillin/streptomycin, 10 mg/mL insulin, 1 mM dexamethasone, 0.5 mM methylxanthine, and 200 mM indomethacin.

Passage 4 human BM-MSCs are used for adipogenic differentiation. Adipogenic differentiation is induced by adipogenic medium.

