Primary CNS neurons used in the manuscript were prepared from forebrains of embryonic day 16 (E16) Sprague-Dawley rats [31 (link)]. Forebrains free of meninges were dissected in ice-cold dissection buffer (Ca2+/Mg2+-free Hank’s Balanced Salt Solution containing 10 mM HEPES) (Invitrogen, Carlsbad, CA, USA), dissociated with L-cysteine activated papain (10 units/mL) (Sigma Aldrich, St. Louis, MO, USA) for 5 min at 37 °C, and resuspended in dissection medium containing trypsin inhibitor (10 mg/mL) (Invitrogen, Carlsbad, CA, USA) for 2–3 min followed by washing with the trypsin inhibitor solution. The tissue was resuspended in a plating medium (NBB27 + glutamate: neurobasal medium containing 2% B27, 1 mM Glutamine, 25 μM glutamic acid, 100 units/mL penicillin, and 100 μg/mL streptomycin) (Invitrogen, Carlsbad, CA, USA) and triturated with a fire-polished glass Pasteur pipette. The cells were then passed through a 70 μm cell sieves and live cells were counted using a hemocytometer and trypan blue exclusion assay.
L cysteine activated papain
Manufactured by Merck Group
Sourced in United States, Germany
L-cysteine activated papain is a laboratory enzyme product. It is an extract from papaya latex that has been activated by the amino acid L-cysteine. The core function of this product is to catalyze the hydrolysis of proteins.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using l cysteine activated papain
1
Isolation and Culture of Rat Primary CNS Neurons
2
Isolation and Culture of Primary Rat Neurons
3
Intra-articular Papain-Induced Osteoarthritis Model
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Following adaptive feeding for 1 week, all rats were randomly divided into control (n=10, regular feeding) and treatment (n=40) groups according to a 50:50 ratio of male:female rats. The rats in the treatment group received a 12 µl intra-articular injection of 1 U/ml L-cysteine-activated-papain in phosphate-buffered saline (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) in the bilateral knee joints at 1, 4 and 7 days to induce OA, as previously described (15 (link)). At 2 weeks after the final injection of papain, the rats in the treatment group were randomly divided into four groups in each group (n=10), namely a model group (regular feeding) and three experimental groups that received a WSDW with the following ratios: WSDW=1:1, WSDW=1:2 and WSDW=2:1. All animals of the treatment groups in the waking state were restrained on an operating platform, and the WSDW machine (XX-hx01LB; Fujian Province Xiangxing Electronic Technology Co., Ltd., Fuzhou China) covered the surface of the knee joint. The treatment groups were then treated for 30 min/day for 12 weeks, after which the animals were sacrificed. The changes in cartilage structure were observed by optical microscopy and transmission electron microscopy (TEM; H-7650, Hitachi, Ltd., Tokyo, Japan).
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