Impact of zonal CHs cocultured with juvenile MSCs (JMSCs) or AMSCs. (A) Zonal CHs (S, M, and D), and JMSCs and AMSCs were expanded and labeled with CellTracker (MSC, green; CH, red). Zonal CHs were mixed with JMSCs or AMSCs (MSC:CH, 4:1). (B and C) Mixed cell populations were well distributed within the constructs with stable ratios through 56 d of culture (B, JMSC/JCH; C, AMSC/JCH). (D) Equilibrium modulus (EY; in kilopascals). (JMSC/JCH, Top; AMSC/JCH, Bottom; light bar, day 28; dark bar, day 56; n = 4/group, *P < 0.05). (E) Efficacy of coculture (solid, AMSC/JCH; hollow, JMSC/JCH; circle, S–M; triangle, M–M; square, D–M).