RGCB-Molecular Platforms: Therapeutic Efficacy Testing Facility

RGCB Molecular Platforms is a state of art facility with well-equipped infrastructure to perform pre-clinical testing of novel molecules for their therapeutic efficacy. We have standardized a series of in vitro and in vivo assays and are in a constant process of adding more assays and making continuous improvement the existing resources. We have a team of excellent scientists with different expertise and always available for consultation. They together with an experienced team of dynamic technologists constitute RGCB Molecular Platforms. Our aim is to support the scientific community for their research needs at a competitive "not for profit" prices while providing top quality of analysis, trouble shooting and data management. Our existing facilities and service offered are listed below.

FLOW CYTOMETRY FACILITY

This core facility provides flow cytometric services and support for the use of flow cytometric techniques. We are equipped with BD FACS Aria II 14 color system and BD FACS Aria III 16 color system. We provide cytometric services required for a vast array of applications.
Services offered

  1. Cell cycle analysis :

    Different cell lines after exposure to test compounds will be stained with propidium iodide and distribution of cells in various phases of cell cycle will be analysed through FACS.

    Cost

    RGCB Users: Rs. 1000 per sample
    Clients from Academic Institutions: Rs. 2000 per sample
    Clients from the Industry or Overseas: Rs.4500 or ($75) per sample
    (One concentration of compound in triplicate/time point/cell)

  2. Cell sorting facility :

    We also provide sterile and non-sterile sorting of various cell types at sorting rates of 20000 cells/second.

    Cost

    RGCB Users: Rs 1000 per hour
    Clients from Academic Institutions: Rs 3000 per hour
    Clients from the Industry or Overseas: Rs.6000 ($100)/hour

  3. Screening for proteasome inhibitors :

    This is a high-throughput image based screening strategy developed to identify specific proteasome inhibitors using stable cells expressing proteasome sensor vector. We have developed two cancer cell lines HCT116 and U20S cells which stably express the proteasome sensor so that lead molecules can be screened either by automated microscopy or by flow cytometry.

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 6000 ($100)
    (Four concentrations of compound in triplicate/time point/cell/sample)

CELLULAR IMAGING FACILITY

This includes facilities for wide field microscopy, Time lapse microscopy, confocal microscopy and spectral confocal microscope with resonance scanner.
Services offered

  1. Apoptosis (Live cell assay) : Both 2D and 3D models :

    Our live cell assay measures the ability of the drug to induce caspase activation in cancer cells using stable cells expressing FRET probe for activated caspases. We have cells engineered with a FRET probe in the nucleus so that automated segmentation and analysis is quite easy to perform using high-throughput imagers
    The following validated cell lines are available with stable expression of FRET probe NLS. (More will be included in the list soon).
    Breast cancer: MCF-7, T47D
    Colon cancer: SW480, HCT 116, SW620
    Cervical Cancer: SiHa, HeLa
    Osteosarcoma: U205
    Neuroblastoma: U251
    Ovarian Cancer: OVCAR8, NCI Adr.Res
    Pancreatic cancer (MiaPaca2, PANC-1)
    (Any relevant three cells may be ideal for initial screening).

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 6000 ($100)
    (Four concentrations of compound in triplicate/time point/cell/sample)

  2. Assays for Autophagy and screening of m-TOR pathway inhibitors :

    Induction of autophagy and inhibitors of m-TOR pathway is also emerging as an important target in cancer. We have generated stable cells expressing LC3 EGFP and its translocation to autophagic vacuoles is used as the readout by imaging approaches and is adapted for high-throughput image based drug screening.
    Available cell lines: OVCAR8, HCT116, DLD

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 6000 ($100)
    (Four concentrations of compound in triplicate/time point/cell/sample)

  3. Mitophagy inducers or inhibitor screening :

    The strategy employs high-throughput image based approach using stable cancer cell lines expressing sensors of Mitophagy.

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 6000 ($100)
    (Four concentrations of compound in triplicate/time point/cell/sample)

  4. Angiogenesis:

    Method 1: We have immortalized human micro- vascular endothelial cells that form quantifiable angiogenic tubules on matrigel. Hence inhibition of tubule formation can be used as the read out for activity. Since the cell line is immortalized, available for screening of large number of compounds using an image based system is feasible.

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 6000 ($100)
    (Four concentrations of compound in triplicate/time point/cell/sample)

    Method 2: Rat aortic ring assay: Dorsal aortic rings will be prepared from rat aorta will be placed in a collagen pre-coated 96-well plates and treated with compounds of interest. Rings will be analysed through phase-contrast microscopy to study microvessel out growth.

    Cost

    RGCB Users:Rs 3000
    Clients from Academic Institutions: Rs 4000
    Clients from the Industry (or Overseas): Rs 9000 ($150)
    (Four concentrations of compound in triplicate/time point/cell/sample)

  5. Assay for antioxidant potential using mammalian cells :

    Nuclear factor erythroid 2-related factor 2 (Nrf2) is the master transcription factor of the antioxidant response element pathway, coordinating the induction of detoxifying and antioxidant enzymes. Nrf2 is normally sequestered in the cytoplasm by Kelch-like ECH-associating protein 1 (Keap1) and upon activation tans ocate to nucleus. We have developed NRF2 EGFP stable cells so that nuclear translocation can be used as a platform for initial screening. An approach for identification of strong antioxidants that break the interaction is also available and utilizes FRET approach.

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 6000 ($100)
    (Four concentrations of compound in triplicate/time point/cell/sample)

  6. Endoplasmic reticulum calcium releasing compounds

    We also developed high throughput calcium imaging assay based on a FRET-based calcium indicator at single cell resolution for compound screening.

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 6000 ($100)
    (Four concentrations of compound in triplicate/time point/cell/sample)

  7. Tumor stem cell assays :

    Tumor stem cells contribute both for tumor initiation and tumor drug resistance forming an ideal target in cancer. However, they are extremely rare in cancer cells and so difficult to be used for drug screening. We have designed several assays to study the effect of compounds on tumor stem cells. From established cell cultures and surgically dissected tumor samples, we have optimized conditions to isolate stem cell like cells by FACS which can be used for identifying compounds that specifically target tumor stem cells using 2D and 3D culture system
    The spheroid formation assay can also be used to study cancer stem cells. Here cells will be plated in low attachment plates and then treated with compounds of interest. Cells will be observed under phase-contrast microscope to analyse spheroid formation. Stem cells will form spheroids under these conditions and effect of compounds on spheroid formation will be analysed.

    Cost

    RGCB Users:Rs 3000
    Clients from Academic Institutions: Rs 4000
    Clients from the Industry (or Overseas): Rs 6000 ($100)
    (Four concentrations of compound in triplicate/time point/cell/sample)

Molecular Screening Facility :

This resource facility is equipped with various resources for screening small molecules for different molecular properties.
Services offered

  1. Quantitative dose-response analysis of growth inhibitory activity or cytotoxicity :

    This will be the preliminary screening where compounds will be tested for their cytotoxicity and optimum doses for further studies will be evaluated.

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 5500 ($90)
    (Four concentrations of compound in triplicate/time point/cell/sample)

  2. Screening for HIF inhibitors :

    The discovery of hypoxia inducible factors (HIFs), which mediate transcriptional responses to changes in oxygen levels will allow this signalling be used for drug screening compounds that inhibit Hypoxic response in cancer. We have developed stable cells with reporter vectors with HRE-Luc or HRE-mCherry-Luc that will allow identification of inhibitors by Luciferase assay.

    Cost

    RGCB Users:Rs 2000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 5500 ($90)
    (Four concentrations of compound in triplicate/time point/cell/sample)

  3. Gene expression studies :

    We have two platforms available. Step One plus (Invitrogen 96-well format) and ABI 7900HT (96 and 384 well format).

    Cost

    RGCB Users:Rs 1000
    Clients from Academic Institutions: Rs 3000
    Clients from the Industry (or Overseas): Rs 5500 ($90)
    (One gene and housekeeping gene)

  4. Next generation sequencing:

    We have equipped with Ion Torrent and PGM platform from Invitrogen (http://rgcb.res.in/genomics-core-facility.php) and capable of doing various assays including exome sequencing, transcriptome sequencing, microbiome analysis etc. We have in house computation facility for analysis. For pricing contact us with your specific requirement.

  5. Microarray analysis:

    We have equipped with Affymetrix platform and help you to address your research needs. For pricing contact us with your specific requirement

Animal Research Facility :

The primary function of Animal Research Facility (ARF) is to assist investigators in their obligation to plan and conduct animal experiments with the highest scientific, humane and ethical principles. This is achieved by development and maintenance of a comprehensive, high quality animal care program, complies with all guidelines and laws concerning with experiments on animals. All studies involving laboratory animals will require prior approval of Institutional Animal Ethics Committee (IAEC) and experiments will strictly follow the rules and regulations laid down by CPCSEA. The animal research facility is equipped with facilities for housing transgenic and immune deficient animals and an in vivo imaging system (IVIS).
Services offered

  1. Angiogenesis :

    Tumor specific capillary formation assay: This is an in vivo assay where tumor cells will be injected intradermally to mouse followed by the administration of compound of interest. At day 10 the skin on ventral side will be collected and tumor directed capillaries will be counted.

    Cost

    RGCB Users:Rs 3000
    Clients from Academic Institutions: Rs 5000
    Clients from the Industry (or Overseas): Rs 11,000 ($180)

    The study will have two groups; control (treated with vehicle; n=5) and treated (one concentration of compound, n=5).

  2. Tumor stem cells (in vivo assays) :

    In vivo tumorigenic potential: Sorted stem cells will be injected to the nude mouse to evaluate its tumorigenic potential.

    Cost

    RGCB Users:Rs 40,000
    Clients from Academic Institutions: Rs 60,000
    Clients from the Industry (or Overseas): Rs 100,000 ($1800)

    The study will have minimum of 3 nude mice/cell type

  3. Pre-clinical tumor models :

    We have developed several in vivo tumor model systems for pre-clinical validation of compounds. Available tumor models are listed below and we are in the process updating the list

    • Human tumor xenografts in athymic nude or SCID mice
    • Orthotopic human tumor models in nude mice (Colon and pancreas)
    • Syngeneic mouse tumor models (Colon and breast)
    • Chemical carcinogenesis models in mice and rats (Colon, gastric, breast and lung)
    Cost

    Studies involving nude mice: n=5 mice/group/tumor model (minimum two groups) :

    RGCB Users: Rs. 50000
    Clients from Academic Institutions: Rs.70000
    Clients from the Industry (or Overseas): Rs.80000 ($1400)


    Cost

    Studies involving nude mice: n=5 mice/group/tumor model (minimum two groups): : with IVIS imaging

    RGCB Users: Rs. 50000
    Clients from Academic Institutions: Rs.70000
    Clients from the Industry (or Overseas): Rs.80000 ($1400)


    Cost

    Studies involving regular mice (BALB/c or C57Bl/6) for syngeneic tumor models: n=5 mice/group/tumor model (minimum two groups) : with IVIS imaging

    RGCB Users:Rs.4000
    Clients from Academic Institutions: Rs.8000
    Clients from the Industry (or Overseas): Rs.10000 ($175)<\p>


    Cost

    Studies involving regular mice (BALB/c or C57Bl/6 or swiss albino) for chemical carcinogenesis models: n=5 mice/group/tumor model (minimum two groups)

    RGCB Users: Rs.5000
    Clients from Academic Institutions: Rs.8000
    Clients from the Industry (or Overseas): Rs.10000 ($175)


    Cost

    Studies involving rats (Wistar or Sprague-Dowly) for chemical carcinogenesis models: n=5 mice/group/tumor model (minimum two groups)

    RGCB Users: Rs.8000
    Clients from Academic Institutions: Rs.12000
    Clients from the Industry (or Overseas): Rs.15000 ($250)

  4. Toxicity profiling (ADME/PK studies)

    This facility is equipped with provision to study toxicity profile of compounds in mice or rat models. Based on client requirement we can conduct acute/sub-acute or chronic toxicity studies.

    Cost

    will be calculated based on the nature of the study, groups involved, choice of the animals (mice or rat) and parameters accessed.

  5. Autoimmune and inflammatory disease models
    • Collagen-induced arthritis model in mice
    • Adjuvant-induced arthritis model in rats
    • Sepsis models in mice
    Cost

    Studies involving regular mice (BALB/c or C57Bl/6 or swiss albino) for arthritis and sepsis models: n=5 mice/group (minimum two groups) :
    RGCB Users:Rs.5000
    Clients from Academic Institutions: Rs.8000
    Clients from the Industry (or Overseas): Rs.10000 ($175)


    Cost

    Studies involving rats for arthritis models: n=5 rats/group (minimum two groups) :
    RGCB Users: Rs.8000
    Clients from Academic Institutions: Rs.10000
    Clients from the Industry (or Overseas): Rs.15000 ($250)

For more information and details, contact:
Dr T.R Santoshkumar (trsanthosh@rgcb.res.in or + 91 09447117380)
or
Dr K.B Harikumar (harikumar@rgcb.res.in or + 91 09567655337)