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Merck
CN
  • Highly Efficient Cardiac Differentiation and Maintenance by Thrombin-Coagulated Fibrin Hydrogels Enriched with Decellularized Porcine Heart Extracellular Matrix.

Highly Efficient Cardiac Differentiation and Maintenance by Thrombin-Coagulated Fibrin Hydrogels Enriched with Decellularized Porcine Heart Extracellular Matrix.

International journal of molecular sciences (2023-02-12)
Fatemeh Navaee, Philippe Renaud, Alexander Kleger, Thomas Braschler
摘要

Biochemical and biophysical properties instruct cardiac tissue morphogenesis. Here, we are reporting on a blend of cardiac decellularized extracellular matrix (dECM) from porcine ventricular tissue and fibrinogen that is suitable for investigations employing an in vitro 3D cardiac cell culture model. Rapid and specific coagulation with thrombin facilitates the gentle inclusion of cells while avoiding sedimentation during formation of the dECM-fibrin composite. Our investigations revealed enhanced cardiogenic differentiation in the H9c2 myoblast cells when using the system in a co-culture with Nor-10 fibroblasts. Further enhancement of differentiation efficiency was achieved by 3D embedding of rat neonatal cardiomyocytes in the 3D system. Calcium imaging and analysis of beating motion both indicate that the dECM-fibrin composite significantly enhances recovery, frequency, synchrony, and the maintenance of spontaneous beating, as compared to various controls including Matrigel, pure fibrin and collagen I as well as a fibrin-collagen I blend.

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Sigma-Aldrich
纤维蛋白原 来源于人类血浆, 50-70% protein (≥80% of protein is clottable)
Sigma-Aldrich
胃蛋白酶 来源于猪胃粘膜, lyophilized powder, ≥3,200 units/mg protein
Sigma-Aldrich
蛋白酶 K 来源于林伯氏白色念球菌, buffered aqueous glycerol solution, Molecular Biology, ≥800 units/mL
Sigma-Aldrich
Thrombin 来源于人类血浆, lyophilized powder, ≥2800 NIH units/mg protein (E1%/280, 18.3)
Sigma-Aldrich
单克隆抗肌钙蛋白 T 小鼠抗, clone JLT-12, purified from hybridoma cell culture