Table 1. Preparation prior to in vitro co-culture experiment start Day 0.

Day Cells/Scaffolds	< -30 days Biologic Preparation	-8 days sMSC transduction	-6 days	-5 days sConstruct preparation	-3 days Chondrocyte preparation	Day 0 Experiment start
sECMs	sECM decellularized from fresh synovium, cut into 6 mm ⊘ discs and frozen (-80°C).			sECM discs thawed at room temperature for 3 hrs and placed in the 6 mm ⊘ insert.		sECM inserts followed alone and placed into plate wells containing chondrocytes
sMSCs	sMSCs prepared from fresh synovium and passaged.	sMSC transduced with Ad-BMP2.	ELISA for BMP2 in the Media from sMSC-BMP2. confirmed transduction and BMP2 greater than sMSC.	106 viable cells in 10% FBS were placed on sECM in insert.
Chondrocytes	Chondrocytes isolated from digestion of fresh cartilage and frozen at -80°C.				Chondrocytes thawed, 103 cells in 10% FBS were placed into each well of the 12 well plate for the experiments assigned to co-culture. Medium was changed daily.	Chondrocytes followed alone and in the plate wells of co-cultures
sConstruct				Inserts were placed into 12 well plates containing 40% FBS. Medium replaced daily, 10% FBS in insert, 40% FBS in well.		sConstruct inserts were placed into plate wells containing chondrocytes.