Priming Adipose-Derived Mesenchymal Stem Cells with Hyaluronan Alters Growth Kinetics and Increases Attachment to Articular Cartilage

Figure 6

Treated MSCs seeded onto articular cartilage explants for 24 hours and three days. MSCs expanded until passage 2 in control media. On the third day flasks were treated with either control media or 1 mg/mL HA media for another three days. Cells were then stripped and cell suspensions were counted using the standard enumeration technique. Cells were seeded into ultra-low adherence 96-well plates plugged with articular cartilage explants in the correct orientation. Conditions (-axis) tested were cells grown in the control flask and seeded in control media (control) (−−), grown in control media and seeded in 1 mg/mL HA media (control in HA) (− +), grown in 1 mg/mL HA media and seeded in control media (primed) (+ −), and grown in 1 mg/mL HA media and seeded in 1 mg/mL HA media (HA) (+ +) and articular cartilage explants alone (no cells seeded to serve as a blank absorbance). Conditions were tested using five technical replicates per 96-well plate and run in biological triplicate (). Wells were assayed at the endpoint using Cell Counting Kit-8 and absorbance of the resulting media alone read at a wavelength of 450 nm (-axis). All conditions were compared back to the control using a -test ( = 0.19, value < 0.05). (a) MSCs seeded for 24 hours (adherence) into ultra-low adherence 96-well plates plugged with articular cartilage explants and (b) MSCs seeded for three days (proliferation) into ultra-low adherence 96-well plates plugged with articular cartilage explants.

(a)

(b)