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.
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.
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.
We also provide sterile and non-sterile sorting of various cell types at sorting rates of 20000 cells/second.
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.
This includes facilities for wide field microscopy, Time lapse
microscopy, confocal microscopy and spectral confocal microscope with
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
Ovarian Cancer: OVCAR8, NCI Adr.Res
Pancreatic cancer (MiaPaca2, PANC-1)
(Any relevant three cells may be ideal for initial screening).
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
The strategy employs high-throughput image based approach using stable cancer cell lines expressing sensors of Mitophagy.
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.
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.
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.
We also developed high throughput calcium imaging assay based on a FRET-based calcium indicator at single cell resolution for compound screening.
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
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.
This resource facility is equipped with various resources for
screening small molecules for different molecular properties.
This will be the preliminary screening where compounds will be tested for their cytotoxicity and optimum doses for further studies will be evaluated.
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.
We have two platforms available. Step One plus (Invitrogen 96-well format) and ABI 7900HT (96 and 384 well format).
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.
We have equipped with Affymetrix platform and help you to address your research needs. For pricing contact us with your specific requirement
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).
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.
In vivo tumorigenic potential: Sorted stem cells will be injected to the nude mouse to evaluate its tumorigenic potential.
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
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.
For more information and details, contact:
Dr T.R Santoshkumar (firstname.lastname@example.org or + 91 09447117380)
Dr K.B Harikumar (email@example.com or + 91 09567655337)