Chondrogenic differentiation of mouse induced pluripotent stem cells using the three‐dimensional culture with ultra‐purified alginate gel

Abstract

As articular cartilages have rarely healed by themselves because of their characteristics of avascularity and low cell density, surgical intervention is ideal for patients with cartilaginous injuries. Because of structural characteristics of the cartilage tissue, a three‐dimensional culture of stem cells in biomaterials is a favorable system on cartilage tissue engineering. Induced pluripotent stem cells (iPSCs) are a new cell source in cartilage tissue engineering for its characteristics of self‐renewal capability and pluripotency. However, the optimal cultivation condition for chondrogenesis of iPSCs is still unknown. Here we show that a novel chondrogenic differentiation method of iPSCs using the combination of three‐dimensional cultivation in ultra‐purified alginate gel (UPAL gel) and multi‐step differentiation via mesenchymal stem cell‐like cells (iPS‐MSCs) could efficiently and specifically differentiate iPSCs into chondrocytes. The iPS‐MSCs in UPAL gel culture sequentially enhanced the expression of chondrogenic marker without the upregulation of that of osteogenic and adipogenic marker and histologically showed homogeneous chondrogenic extracellular matrix formation. Our results suggest that the pluripotency of iPSCs can be controlled when iPSCs are differentiated into iPS‐MSCs before embedding in UPAL gel. These results lead to the establishment of an efficient three‐dimensional system to engineer artificial cartilage tissue from iPSCs for cartilage regeneration. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1086–1093, 2019.

Shaking culture enhances chondrogenic differentiation of mouse induced pluripotent stem cell constructs

“…To date, several studies have demonstrated induction of chondrogenic differentiation of mouse iPSCs; however, the induction methods used in previous reports are slow and require a long period of time. For example, Hontani et al 6 reported a method to induce chondrogenic differentiation of mouse iPSCs using 3D culture with an ultra-purified alginate gel, in which iPSCs were first guided to differentiate into MSCs by repeated passaging. Then, the cells were seeded on the alginate scaffold and subjected to chondrogenic differentiation for 28 days.…”

“…For example, Hontani et al . 6 reported a method to induce chondrogenic differentiation of mouse iPSCs using 3D culture with an ultra-purified alginate gel, in which iPSCs were first guided to differentiate into MSCs by repeated passaging. Then, the cells were seeded on the alginate scaffold and subjected to chondrogenic differentiation for 28 days.…”

“…However, to date there are few published studies regarding therapeutic applications of iPSCs in cartilage diseases, possibly because robust and reproducible protocols to fabricate cartilaginous tissues from iPSCs have not yet been established 5 . Some methods have been reported to achieve chondrogenic induction of 3D cultures of iPSCs; however, these methods are time-consuming and require complicated procedures 3 , 6 , 7 . Therefore, chondrogenic differentiation of iPSCs awaits effective protocols to rapidly guide the differentiation of the iPSCs toward chondrogenic cells or tissue.…”

Mechanotransducive Biomimetic Systems for Chondrogenic Differentiation In Vitro

“…Embryonic stem cells seeded on elastic polydimethylsiloxane scaffolds and subjected to mechanical loading resulted in robust induction of chondrogenic differentiation [ 30 ]. Embryonic stem cell application-related ethical issues can be avoided by replacement with iPSCs that were also shown to be effective in chondrogenic ECM production [ 31 ]. However, the difficulties in directing pluripotent stem cells towards specific mature lineages and iPSCs genomic instability often complicates their use in different applications [ 32 ].…”

iPSCs in Modeling and Therapy of Osteoarthritis

“…Until today, there have been only a few published attempts at combing biomaterials with iPSCs to promote chondrogenic differentiation. For instance, Hontani et al [ 38 ] came up with a novel chondrogenic differentiation method based on a combination of 3D cultivation in ultra-purified alginate gel as a scaffold and step-wise differentiation of iPSCs into chondrocytes via the MSC-like cell stage. iPSC-MSCs in alginate gel were supplemented with MSC induction medium.…”