Application of recombinant human PH20 to preparation of medicines used for treating thyroid-associated oculopathy
A technology for thyroid and eye diseases, applied in the field of biopharmaceuticals
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Embodiment 1
[0109] 1. Materials and methods
[0110] (1) Reagents and antibodies
[0111] High glucose DMEM medium (Gibco 11965092), recombinant human PH20 (Abcam ab132258), recombinant human IGF-1 (R&D 291-G1-200), recombinant human TGF-β1 (PeproTech 100-21), rabbit anti-human α-SMA (Abcam ab5694), mouse anti-human Vimentin (Abcam ab8978), donkey anti-rabbit-488 (Invitrogen A21206), donkey anti-mouse Cy3 (Jackson ImmunoResearch, 715165150), TRIZOL (Life 135306), reverse transcription kit (TAKARARR047A ), fluorescent quantitative PCR kit (TAKARA RR420A), high-fidelity PCR enzyme (TAKARA R040A), hyaluronic acid detection kit (Cloud-Clone CEA182Ge), type I collagen detection kit (Abcam ab210966), BrdU detection kit ( Roche 11647229001)
[0112] (2) Primary culture of fibroblasts
[0113] During orbital decompression, 5*5*5mm of muscle tissue was cut from the patient’s extraocular muscle belly 3 , and then immediately put it into the culture medium without FBS, and store it in the refrig...
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