For efficient delivery of siRNA into liver metastasis, lipoplexes with 50 µg Cy5.5-siRNA were administered intravenously into mice with HeLa-liver metastasis at 1 min after the intravenous injection of 1 mg chondroitin sulfate (Wako Pure Chemical Industries, Ltd.) at 10 days after inoculation of HeLa-Luc cells, as reported previously (sequential injection method) (15 (link),16 (link)). A total of 1 h after injection, the mice were sacrificed, and Cy5.5 fluorescent imaging of the tissues was performed using a NightOWL LB981 NC100 system (Berthold Technologies, Bad Wildbad, Germany). In Cy5.5 fluorescent imaging, the excitation and emission filters were set at 630/20 and 680/30 nm, respectively. The exposure time for fluorescence was 5 sec. A grayscale body-surface reference image was collected using a NightOWL LB981 CCD camera (Berthold Technologies). The images were analyzed using IndiGo2 software (version 2.0.1.0; Berthold Technologies) provided with the in vivo imaging system.
Chondroitin sulfate
Manufactured by Fujifilm
Sourced in Japan
Chondroitin sulfate is a chemical compound found in the cartilage and connective tissues of the human body. It is commonly used in laboratory settings for various research and analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Collagenase a, by Roche (3 mentions)
Opti mem 1, by Thermo Fisher Scientific (3 mentions)
Ascorbic acid, by Merck Group (3 mentions)
103 tryple select, by Thermo Fisher Scientific (2 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions)
Calcium chloride, by Merck Group (2 mentions)
Type 1 collagen coated six well plate, by Corning (2 mentions)
Nightowl lb981 nc100 system, by Berthold Technologies (1 mentions)
Indigo2, by Berthold Technologies (1 mentions)
Sb431542, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Calcium chloride, by Fujifilm (1 mentions)
5 protocols using chondroitin sulfate
1
In Vivo Imaging of Liver Metastasis
2
Collagen-Chondroitin Sulfate Hydrogel Fabrication
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A 1% collagen solution was prepared by dissolving 0.1 g of lyophilized collagen (rHCI and rHCIII, from Fibrogen) in 10 ml of ultra-pure ddH2O. Chondroitin sulfate (CS; Wako), N-ethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) were added to produce a final mixture with a mass ratio of 1:4:0.5:0.3 for collagen:CS:NHS:EDC. The materials were prepared on ice using an enclosed system that allows homogenous mixing without adding bubbles. After thorough mixing, the pH was adjusted to 7.4 by NaOH (1.0 N). Matrices without CS were prepared similarly but with PBS added to compensate for the volume of CS. Matrices labeled with Alexa-Fluor®594-NHS were prepared by adding 20 µL of the dye stock solution (1 mg/mL in DMSO) to the gels before adding the NaOH (25 nmol of dye per hydrogel), followed by 20 mixing steps.
3
Human Corneal Endothelial Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Four human donor corneas (from persons >40 years of age) were used for human CEC (HCEC) cultivation, as described previously [17 (link)]. Briefly, Descemet’s membranes containing the HCECs were stripped from the corneas and the membranes were incubated with 1 mg/mL collagenase A (Roche Applied Science) at 37°C for 12 hours. The HCECs were then seeded in one well of a 48-well plate coated with laminin E8 fragments (iMatrix-511; Nippi, Incorporated, Tokyo, Japan) (2.0 μg/cm2) [17 (link)] [17 (link)]. The culture medium was prepared according to published protocols. First, basal medium was prepared, consisting of OptiMEM-I (Life Technologies Corp., Carlsbad, CA), 8% FBS, 5 ng/mL epidermal growth factor (Sigma-Aldrich Co., St. Louis, MO), 20 μg/mL ascorbic acid (Sigma-Aldrich Co.), 200 mg/L calcium chloride, 0.08% chondroitin sulfate (Wako Pure Chemical Industries, Ltd., Osaka, Japan), 50 μg/mL gentamicin, and 10μM SB431542 (Merck Millipore, Billerica, MA). This basal medium was then conditioned by culturing human bone marrow-derived mesenchymal stem cells (BM-MSCs) for 24 hours. Finally, the conditioned basal medium was collected for use as the culture medium for HCECs. Cultivated HCECs were used at passages 3 through 6 for the experiments.
4
Culturing Primary Human Corneal Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Unless otherwise stated, the HCECs were cultured according to the published protocols, with some modifications. 11 Briefly, the Descemet's membranes with the CECs were stripped from donor corneas and digested at 378C with 1 mg/mL collagenase A (Roche Applied Science, Penzberg, Germany) for 2 hours. The HCECs obtained from a single donor cornea were seeded in one well of a Type-I collagen-coated six-well plate (Corning, Inc., Corning, NY, USA). The culture medium was prepared according to published protocols. Briefly, basal medium was prepared with OptiMEM I (Life Technologies Corporation, Carlsbad, CA, USA), 8% fetal bovine serum (FBS), 5 ng/mL epidermal growth factor (EGF; Life Technologies), 20 lg/mL ascorbic acid (Sigma-Aldrich Corp., St. Louis, MO, USA), 200 mg/L calcium chloride (Sigma-Aldrich Corp.), 0.08% chondroitin sulfate (Wako Pure Chemical Industries, Ltd., Osaka, Japan), and 50 lg/mL gentamicin. Mesenchymal stem cell (MSC)conditioned medium was prepared as previously described. 18 The HCECs were cultured using MSC-conditioned medium at 378C in a humidified atmosphere containing 5% CO 2 , and the culture medium was changed twice per week. The HCECs were passaged at ratios of 1:3 using 103 TrypLE Select (Life Technologies) at 378C for 12 minutes when they reached confluence. The HCECs at passages two through five were used for all experiments.
5
Culturing Human Corneal Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HCECs obtained from 30 human donor corneas at distinct ages were cultured according to published protocols, with some modifications. 13 Briefly, the Descemet's membranes with the CECs were stripped from donor corneas and digested at 378C with 1 mg/mL collagenase A (Roche Applied Science, Penzberg, Germany) for 2 hours. The HCECs obtained from a single donor cornea were seeded in one well of a Type-I collagen-coated six-well plate (Corning, Corning, NY, USA). The culture medium was prepared according to published protocols. 13 Briefly, basal medium was prepared with Opti-MEM-I (Life Technologies Corporation, Carlsbad, CA, USA), 8% fetal bovine serum (FBS), 5 ng/ mL epidermal growth factor (Life Technologies), 20 lg/mL ascorbic acid (Sigma-Aldrich, St. Louis, MO, USA), 200 mg/L calcium chloride (Sigma-Aldrich), 0.08% chondroitin sulfate (Wako Pure Chemical Industries, Osaka, Japan), and 50 lg/mL gentamicin. The HCECs were cultured at 378C in a humidified atmosphere containing 5% CO 2 , and the culture medium was changed twice per week. When they reached confluence, the HCECs were passaged at ratios of 1:3, after the treatment with 103 TrypLE Select (Life Technologies) for 12 minutes at 378C. The HCECs at passages two through five were used for all experiments.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!