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CHONDRO-RT-PCR real time

Quantification assay for chondrogenic differentiation of MSCs         cod. BM-015

Principle of the test: Quantitative analysis of ACAN mRNA expression

Technology: Relative Quantitative Real Time PCR

Gene Target: ACAN

Specimen: cDNA

Results: ΔΔCt method

Reporting Units: Arbitrary Units (AU)

Number of tests: 25 tests BM-015 

Kit storage: -20°C

Necessary equipment: Thermocycler, 7500 Real Time PCR System

Status: Ready to use

CHONDRO-RT-PCR real time  cod. BM-015

Quantification assay for chondrogenic differentiation of MSCs

·      CONDRO-RT-PCR real time Quantification complete kit 25 tests BM-015

Adult human mesenchymal stem cells (MSCs) attracted the most attention for cartilage tissue engineering studies, because of their high proliferation rate, easy availability and capacity to differentiate into multiple cell types34. The precise predictions of the differentiation direction and potential of MSCs are an important key to the success of regenerative medicine. MSCs can differentiate into various cell types, including osteoblasts, chondrocytes, or adipocytes; therefore, they are promising as regenerative medicine. The chondrogenic differentiation of MSCs has been well demonstrated in vitro, provided by the use of inductive media containing differentiation-promoting agents and growth factors35. During the process of chondrogenic differentiation of MSCs, Sox9 collaborates with Sox5 and Sox6 to regulate the expression of Col2a1, IX, XI, and Acan, specific markers of cartilage36. The ACAN gene provides instructions for making the aggrecan protein. Aggrecan is a type of protein known as a proteoglycan, which means it has several sugar molecules attached to it. It is the most abundant proteoglycan in cartilage, a tough, flexible tissue that makes up much of the skeleton during early development. Most cartilage is later converted to bone (a process called ossification), except for the cartilage that continues to cover and protect the ends of bones and is present in the nose, airways, and external ears.

We have established a novel quantitative analysis of Acan mRNA expression. This method is based on real-time PCR. The expression levels of the mRNAs were determined from the threshold cycle (Ct), and the relative expression levels were calculated using the 2-ΔΔCt method. For mRNA quantification, the Ct values were normalized to the expression of the GAPDH mRNA level. Results are expressed in corresponding arbitrary units (AU) User friendly and complete, the CHONDRO-RT-PCR real time Quantification kit is suitable for any laboratory.

Reference

1.      Caplan AI. Adult mesenchymal stem cells for tissue engineering versus regenerative medicine. J Cell Physiol 2007; 213: 341–347.

2.      J. Kim, B. Lin, S. Kim, B. Choi, D. Evseenko and M. Lee, J. Biol. Eng., 2015, 9, 1–11.

3.      F. Long and D. M. Ornitz, Cold Spring Harbor Perspect. Biol., 2013, 5, a008334.