* Description
Genosco provides the tools for high throughput screening of a large number of chemical compounds rapidly and in parallel to determine which is related to osteoclastic activities, and then analyzes the results and chooses further compounds based on this information.
* Advantages
Saves you time of Drug Discovery for the prevention of the osteoporosis Rapid results compared to other target validation technologies Selects for a large number of hit compounds that are effective to inhibit the osteoclastic activities. Optimizes cellular delivery methods and reagents Genosco combines its years of expertise in the field of osteoclastic research technology to create a new option for our clients
* HTS for direct-acting modulators of osteoclastogenesis Direct-acting...OAAS™
Effects of (A) IL-1β, (B) calcitonin, and (C) unknown sample on the activity of osteoclasts in the presence of ODF and M-CSF. Resorption activity is measured as resorbed area.
* Procedure
1. Mouse bone marrow cell culture
Mice were sacrificed by cervical dislocation.     The femur and tibia were removed aseptically, freed of adherent soft tissue, and cut across the epiphysis.     The marrow cavity was flushed. The collected tissues were digested. After digestion, the released cells were treated to remove the red blood cells. After washing, the marrow cells were filtered for single cell dispersion.     The filtered cells were seeded in α-MEM containing FBS, and M-CSF.     After a 24-hour culture, the non-adherent cells were collected and seeded at 96-well OAAS™ plate under α-MEM containing FBS, M-CSF and TGF-β. The cultures were maintained for up to 8-9 days in the presence of M-CSF, TGF-β and ODF.       culturing
2. Screening process
2.1 Measurement of TRAP (+) multinucleated cells After culturing, the cells were rinsed with phosphate buffered saline, fixed with citrate-acetone-formaldehyde and stained for TRAP by incubating the cells with a TRAP staining kit (Sigma). After staining, the TRAP (+) multinucleated cells containing three or more nuclei were counted as osteoclasts using optical microscopy.   2.2. Measurement of resorption pit The mouse bone marrow cells were cultured on a calcium-phosphate coated OAAS™ plate. The culture medium was removed to measure the resorption area, and the cells were detached by treating the culture plates with 5% sodium hypochlorite. The resorption areas were captured using a microscope equipped with a digital camera. The total resorption pit areas of the well surface were measured by image analyzing software, Image pro plusTM (Media Cybernetics Inc., USA).
* Assay results
What sorts of results - Raw data - TRAP test Result: Inhibition percentage will be compared to control. - Resorption Test Results: Selected compound after TRAP will be tested for the inhibition percentage of inhibition and will be compared to the control. The test result will be described with table. - Tested tissue culture plates and OAAS™ plates will be provided on customer’s request - Further service for HIT compound may be discussed. Imaging file will be provided on customer’s request. Sensitivity The grade shows the rate of inhibition of osteoclastic differentiation or osteoclastic activity when compared to control. Grade 0: 0% Grade 1: 20% Grade 2: 40% Grade 3: 60% Grade 4: 80% Grade 5: 100% Grading for TRAP+ 0 1 2 3 4 5 Grading for Resorption Pit 0 1 2 3 4 5 Quantitation or Dose-dependent Response Test is available for HIT compound.
* Capacity
Genosco can initiate approximately 800 assays per day for differentiation or resorption assay. Any screening much larger than 10,000 compounds will take a month or two months.
* Costs
Cost of the contract assay for osteoclast depends on the complexity and size of the contract. Contract assay of less than 80 points may be delayed due to pooling. Screening more than 10,000 compounds will be significantly cheaper per assay. Please contact Genosco to obtain specific advises and estimated price quote for the contract. info@genosco.com