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. 2019 Dec 17;7:411. doi: 10.3389/fbioe.2019.00411

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Copyright © 2019 Lin, Li, Li, Li, Del Duke, Shen, Hao, O'Donnell, Bunnell, Goodman, Alexander, Tuan and Lin.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Fabrication of osteochondral (OC) or chondral (CH) tissue in the dual-flow bioreactor. (A) Schematic of generating OC tissue. After naïve iMPCs were encapsulated into mGL and placed into bioreactor, optimized chondrogenic, and osteogenic media were perfused through the top and bottom flow, respectively, to Induce the formation of the bi-phasic OC tissue, with cartilage in the top and bone in the bottom. (B,C) Characterization of the engineered OC construct. (B) Expression levels of chondrogenic (AGAN, COL2) and osteogenic (OCN, BSP2) marker genes in the top and bottom (Bot) parts of OC construct. Gene expression levels in the top component are normalized as 1. N = 3 per group; *p < 0.05; **p < 0.01. (C) Alcian Blue and Alizarin red staining for OC construct. Bar, top panel = 500 μm; Bar, bottom panel = 200 μm. (D) Schematic of generating CH tissue. All the culture conditions were the same to those used in generating OC, except that no cells were encapsulated in the bottom part. (E,F) Characterization of the engineered CH construct. (E) Expression levels of ACAN, COL2 in the constructs after 28 days differentiation. Expression levels of Day 0 are set as 1. N = 3 per group; ***p < 0.001. (F) Alcian Blue and Alizarin red staining for CH construct. Bar = 500 μm.