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COL2A1Collagen, Type II, Alpha1
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The following cDNAs were used as templates for synthesizing the antisense probes: 0.5 kb Col2a1 [46]; 0.7 kb Matn4 (IMAGE Clone ID: 1366191); 4.7 kb Hapln1 (IMAGE Clone ID: 30430221); 0.5 kb 3110079Q15Rik (IMAGE Clone ID: 40104070); 0.6 kb Sox5 (IMAGE Clone ID: 40047865); and 0.5 kb Sox9 (Sox9 exon 1 sequence from BAC clone RP24-248D4).
As shown in Figure 3, T[beta]RIII RNAi had no obvious effects on GAG and COL II secretion (Figures 3(a) and 3(b); P > 0.05) or any noticeable effects on the expression of cartilage-specific genes (SOX9 and COL2A1) when hMSCs were cultured in control medium (Figure 3(c); P > 0.05).
At day 7, the mRNA expression levels of SOX9, ACAN, and col2a1 significantly increased compared with the negative control group (Figure 4(i)) (P < 0.01).
Both methods have relevance to reproduce changes in phenotype biomarker expression reported in OA chondrocytes [32] including aggrecan, COL2A1, COL2B, MMP-9, and MMP-13, which were evaluated by RT-qPCR.
Likewise, just like N-cadherin, the SOX trio acts together to increase the expression of proteins related to extracellular matrix synthesis, such as COL2A1 [34], the main collagen of cartilage tissue.
Our results showed that FGF-18 promotes chondrogenic differentiation of ADSCs by increasing the expression of SOX-9 and also promotes the expression of chondrocytic matrix gene Col2a1 and aggrecan during chondrogenesis of ADSCs.
Aggrecan (ACAN) and collagen type II (COL2A1) are the major proteoglycan proteins in the articular cartilage that provides the hydrated gel structure.
Chondrogenic markers ACAN and COL2A1 were significantly increased under chondrogenic-inducing conditions in both P2 and P15 hPDLSCs derived from donors (Figure 5(b), A3 and B3, resp.) and MS patients (Figure 5(d), C3 and D3, resp.).
Expression levels of all chondrogenic marker genes were significantly upregulated in MSCs at ELSC following 21 days in chondrogenic media, while ASCs at ELSC demonstrated no change in aggrecan expression following pellet culture, but significant increases in SOX-9, COL2A1, and COL10A1.
The mRNA expression levels for type II collagen alpha1 (Col2a1; Hs 00264051_m1) and Sox9 (Hs1001343_g1) genes were assessed by TaqMan real-time polymerase chain reaction (qRT-PCR, Applied Biosystems, Carlsbad, CA) using a StepOnePlus Real-Time PCR instrument (Applied Biosystems).
Six additional cases of SEDC due to the same and recurrent R989C mutation in the COL2A1 gene-the clinical and radiological follow-up.