Some reports suggest low immunogenicity of chondrocytes 6, 7, whereas others show that chondrocytes are antigenic and elicit varying degrees of immune reactions 8, 9.
However, whether the transplanted allogeneic cartilage causes an immune reaction remains controversial. Allogeneic cartilage has been transplanted clinically without matching human leukocyte antigen (HLA) types and without the use of immunosuppressive drugs 3, 4, 5. Although cell-based therapies have been proposed, only a limited number of autologous chondrocytes are generated since expansion culture steers the chondrocyte character toward that of fibroblastic cells 1, as evidenced in autologous chondrocyte implantation (ACI), wherein more than 90% of the repaired tissue is fibrocartilaginous 2. Cartilage ECM consists of collagen fibrils composed of type II, IX, and XI collagen molecules and proteoglycans composed of aggrecan, link protein, and glycosaminoglycans.Īs articular cartilage has a limited capacity for repair, and so far, no drugs are available for cartilage repair, focal damage or erosion of articular cartilage frequently leads to debilitating conditions, such as osteoarthritis. Articular cartilage is avascular and consists of chondrocytes embedded in the extracellular matrix (ECM), which enables the mechanical functions necessary for joint motion and shock absorption. Our study outcomes suggest that allogeneic transplantation of iPSC-derived cartilage organoids may be clinically applicable for the treatment of patients with chondral defects of the articular cartilage however further assessment of functional recovery long term after load bearing injuries is required.Īrticular cartilage covers the ends of bones and provides lubrication, which is vital for smooth joint movement and shock absorption. Pathway analysis suggested the involvement of SIK3 inactivation. Single-cell RNA-sequence analysis indicated that iPSC-derived cartilage organoids differentiated after transplantation, acquiring expression of PRG4 crucial for joint lubrication. iPSC-derived cartilage organoids integrated with the host native articular cartilage and prevented degeneration of the surrounding cartilage. Histological analysis revealed that allogeneic iPSC-derived cartilage organoids in chondral defects elicited no immune reaction and directly contributed to tissue repair for at least four months.
Here, we show that allogeneic iPSC-derived cartilage organoids survive and integrate as well as are remodeled as articular cartilage in a primate model of chondral defects in the knee joints. However, to the best of our knowledge, allogeneic cartilage transplantation into primate models has never been assessed. Validating custom primers for sequencing on the iSeq 100 Sequencing System.Induced pluripotent stem cells (iPSCs) are a promising resource for allogeneic cartilage transplantation to treat articular cartilage defects that do not heal spontaneously and often progress to debilitating conditions, such as osteoarthritis. Illumina cannot guarantee the performance or compatibility of custom primers. Refer to VP10 and VP14 Custom Primers for more information. Each primer is loaded into the library reservoir individually.ĭepending on your library preparation kit, you might be required to use Illumina primer mixes. The iSeq 100 allows the use of two custom read primers and two custom index primer combinations. The iSeq 100 Sequencing System support pages on the Illumina support center provides additional resources.
#Sequencher illumina primer design install#
To use custom primers with the iSeq 100 i1 Reagent v2 kits, update and install iSeq 100 Control Software v3.0 or later. Prepare and add the appropriate volume of each custom primer to the library reservoir of the cartridge after pre-run checks are completed. Select the custom primers in the control software during run setup.
#Sequencher illumina primer design manual#
Set the iSeq 100 to manual run mode and enable the custom primers setting. Using custom primers for a run on an Illumina iSeq 100 Sequencing System requires three additional steps at the run setup: Follow the run setup instructions described in the Run Settings.
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