Three-dimensional culture and chondrogenic differentiation of mesenchymal stem cells in interconnected collagen scaffolds

Abstract

Interconnected scaffolds are useful for promoting the chondrogenic differentiation of stem cells. Collagen scaffolds with interconnected pore structures were fabricated with poly(lactic acid-co-glycolic acid) (PLGA) sponge templates. The PLGA-templated collagen scaffolds were used to culture human bone marrow-derived mesenchymal stem cells (hMSCs) to investigate their promotive effect on the chondrogenic differentiation of hMSCs. The cells adhered to the scaffolds with a homogeneous distribution and proliferated with culture time. The expression of chondrogenesis-related genes was upregulated, and abundant cartilaginous matrices were detected. After subcutaneous implantation, the PLGA-templated collagen scaffolds further enhanced the production of cartilaginous matrices and the mechanical properties of the implants. The good interconnectivity of the PLGA-templated collagen scaffolds promoted chondrogenic differentiation. In particular, the collagen scaffolds prepared with large pore-bearing PLGA sponge templates showed the highest promotive effect.

Injectable hypoxia-preconditioned cartilage progenitor cells-laden GelMA microspheres system for enhanced osteoarthritis treatment

“…However, 2D culture cannot simulate the practical physiological conditions of stem cells niche and this severely limits the therapeutic efficacy and impedes the clinical translation of stem cells-based therapy. 3D culture system can mimic a more actual cellular physiological condition through the elaborate interactions of cell to cell and cell to extracellular matrix [ 17 , 22 , 23 ]. In addition, previous studies have indicated that 3D cultured MSCs possessed enhanced stemness and improved therapeutic effects [ 22 , 23 ].…”

“…3D culture system can mimic a more actual cellular physiological condition through the elaborate interactions of cell to cell and cell to extracellular matrix [ 17 , 22 , 23 ]. In addition, previous studies have indicated that 3D cultured MSCs possessed enhanced stemness and improved therapeutic effects [ 22 , 23 ]. Therefore, it is instructive to maintain the physiological features of CPCs and maximize the efficacy of CPCs by using 3D culture system.…”

“…Three-dimensional (3D) culture technology can mimic a more actual and beneficial physiological environment through cell to cell and cell to matrix interaction [ [19] , [20] , [21] ]. In addition, previous studies have demonstrated that mesenchymal stem cells (MSCs) cultured in 3D condition possess an enhanced stemness and improved therapeutic effects [ 14 , 22 ]. Previous study showed that 3D cultured bone marrow-derived MSCs in the poly(lactic acid- co -glycolic acid) (PLGA)-templated collagen scaffolds significantly promoted the expressions of chondrogenesis-related genes and the production of cartilage matrix [ 22 ].…”

“…In addition, previous studies have demonstrated that mesenchymal stem cells (MSCs) cultured in 3D condition possess an enhanced stemness and improved therapeutic effects [ 14 , 22 ]. Previous study showed that 3D cultured bone marrow-derived MSCs in the poly(lactic acid- co -glycolic acid) (PLGA)-templated collagen scaffolds significantly promoted the expressions of chondrogenesis-related genes and the production of cartilage matrix [ 22 ]. Additionally, 3D cultured equine bone marrow-derived MSCs as spheroids possess higher amounts of anti-inflammatory and anti-apoptotic factors than 2D cultured MSCs, and thus enhance their anti-inflammatory property [ 23 ].…”

Stepwise Proliferation and Chondrogenic Differentiation of Mesenchymal Stem Cells in Collagen Sponges under Different Microenvironments

“…After reverse transcription with a high-capacity cDNA reverse transcription kit, amplification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH, a housekeeping gene), collagen I (Col1a2), collagen II (Col2a1), aggrecan (Acan), and SOX9 was conducted with a QuantStudio ® 3 Real-Time PCR System (Thermo Fisher Scientific). The previously reported primer and probe sequences as shown below were used [ 52 , 53 ]. GAPDH: (forward) 5′-ATGGGGAAGGTGAAGGTCG-3′, (reverse) 5′-TAAAAGCAGCCCTGGTGACC-3′, (probe) 5′-CGCCCAATACGACCAAATCCGTTGAC-3′; Col1a2: (forward) 5′-CAGCCGCTTCACCTACAGC-3′, (reverse): 5′-TTTTGTATTCAATCACTGTCTTGCC-3′, (probe): 5′−CCGGTGTGACTCGTGCAGCCATC-3′; Col2a1: (forward) 5′GGCAATAGCAGGTTCACGTACA-3′, (reverse) 5′-CGATAACAGTCTTGCCCCACTT-3′, (probe) 5′−CCGGTATGTTTCGTGCAGCCATCCT-3′; Acan: (forward) 5′-TCGAGGACAGCGAGGCC-3′, (reverse) 5′-TCGAGGGTGTAGCGTGTAGAGA-3′, (probe) 5′-ATGGAACACGATGCCTTTCACCACGA-3′; SOX9: (forward) 5′-CACACAGCTCACTCGACCTTG-3′, (reverse) 5′-TTCGGTTATTTTTAGGATCATCTCG-3′, (probe) 5′-CCCACGAAGGGCGACGATGG-3′.…”