Highly motivated, hard-working and diligent post-doctoral research fellow with 7+ years of experience in cell culture and 4+ years in stem cell biology with a goal to advance career in the field of regenerative medicine.
Expertise in cellular reprogramming, hPSC culture and differentiation.
Skilled at 2D and 3D differentiation, especially retinal and cerebral organoid generation for transplant studies.
Developed cGMP grade iPSCs from diseased fibroblasts, followed by progenitor differentiation for cell therapy- based applications.
Worked on dynamic collaborative projects and demonstrated ability at independent planning, execution and analysis of experiments.
The Johns Hopkins University School of Medicine | Baltimore, MDPost-Doctoral Research Fellow05/2018 - Current
Post-doctoral research focuses on generation of transplantable retinal tissue from hPSCs, an extension of one of my graduate research project.
Culture, maintain and characterize several human pluripotent stem cell (hPSC) lines including cord-blood derived iPSCs (CB-iPSC), fibroblast derived iPSCs (F-iPSC) and embryonic stem cells (ESC) in feeder-free conditions.
Revert primed hPSCs to a naïve/ ground state in feeder-dependent and feeder-free conditions and characterization of naïve hPSCs.
Differentiate primed and naïve hPSCs to neural lineage (2D), characterization by immunofluorescence and analysis of kinetics of differentiation by Flow Cytometry.
Differentiate primed and naïve hPSCs to retinal organoids and retinal pigment epithelium (RPE) (3D).
Characterize early and late retinal differentiation phases by qPCR and IF for analysis of differentiation kinetics of primed vs naïve hPSCs.
Dissect neural retinal domains and forebrain neurospheres for generation of retinal cups and optic vesicles respectively, using different protocols followed by long term suspension culture, involving selection and grooming in suspension.
Characterize all six retinal cell types, including photoreceptors, generated sequentially at developmental timepoints by immunofluorescence (cryosection and whole mount) and confocal microscopy imaging.
Generate cerebral organoids from naïve and primed hPSCs (3D), long-term culture and characterization by immunofluorescence (cryosection).
Generation of hPSC reporter line for tracking retinal differentiation using CRISPR/Cas9 gene editing technology.
The Johns Hopkins University School of Medicine | Baltimore, MDVisiting graduate research assistant01/2017 - 04/2018
Generate non-integrated, xeno-free iPSCs from Hutchinson Gilford Progeria Syndrome (HGPS) patient fibroblasts using episomal transcription factors followed by feeder free, xeno-free culture and maintainance.
Characterize HGPS-iPSCs by qPCR, immunofluorescence, flow cytometry, sanger sequencing, teratoma formation and tri-lineage differentiation (EB and monolayer).
Revert HGPS-iPSCs to a naïve/ ground state and characterization of naïve HGPS-iPSCs by qPCR, immunofluorescence, western blot and flow cytometry.
Generate HGPS traffic light reporter lines using lentivirus to study gene correction efficiencies in primed and naïve states.
Design and delivery of CRISPR/Cas9 gene editing system components for gene correction of HGPS iPSCs.
Differentiate HGPS iPSCs, naïve and primed, to mesodermal lineages including vascular progenitors, endothelial cells, pericytes, osteocytes, chondrocytes, adipocytes and vascular smooth muscle cels along with their characterization using flow cytometry, IF, proliferation and senescence detection assays.
Howard University College of Medicine | Washington, DCGraduate Research Assistant10/2013 - 12/2016
Generated iPSCs from HGPS patient fibroblasts using lentiviral, episomal and mRNA- based (NM-RNA and mRNA based) methods via nucleofection and lipid-based delivery in feeder-free and feeder-dependent conditions
Feeder-free culture and maintainance of HGPS iPSCs, characterization by immunofluoroscence.
Genome-wide analysis of DNA methylation changes in Human Embryonic Kidney (HEK) cells treated with Citalopram.
Genome-wide analysis of DNA methylation changes in Hela cells treated with Cannabinoid receptor agonist WIN 55,212-2 mesylate.
Genome-wide analysis of miRNA expression changes in ARPE-19 cells treated with Astaxanthin & Luperox.
Cloning and overexpression of Lamin A and Progerin (mutated Lamin A) encoding vectors in HEK cells through PEI mediated transfection.
Intrexon | San Diego, CAResearch Intern06/2015 - 08/2015
mRNA based reprogramming of human chondrocytes and BJ fibroblasts.
Culture, cryopreservation and characterization of iPSCs by qPCR, immunofluorescence, tri-lineage differentiation through EB formation, followed by characterization of differentiated cell types.
Bovine Egg Precursor cell (DDX4+) isolation from ovarian tissue, In vitro aggregate formation (cumulus oophorus complex) and its differentiation to oocyte-like cell.
RNA isolation and cDNA production from aggregates for determining gene expression of oocyte-like cell.
Project focused on the assessment of neuronal oxidative stress in murine model of rabies.
Infecting swiss albino neonatal mice with rabies virus, harvesting brain tissue, detection of the infection by IF and Nested RT-PCR.
Measured oxidative stress (OS) generated during the course of infection using biochemical assays for protein and reactive-oxygen species detection followed by microplane spectrophotometric analysis.
Cell culture and maintenance of 3T3 cell line.
Sai Pathologyy Clinic | Nasik, MH, IndiaUndergraduate Researcher07/2009 - 03/2010
Project focused on phage therapy development for treating multidrug resistant Infections.
Studying antibiotic profile of multi-drug resistant bacteria (superbugs), isolation of superbugs from sewage water & antibiotic profiling using Kirby-Bauer method.
Isolation of bacteriophages specific to these host organisms through plaque formation, purification of bacteriophage lysates for its downstream application in treatment of drug-resistant nosocomial infections.
iPSC derivation and culture- reprogramming using lentiviral, episomal and mRNA based methods (lipid-based transfection and nucleofection) in feeder-free and feeder-dependent conditions, human pluripotent stem cell (iPSC & hESC) culture (antibiotic free), maintenance (feeder free and xeno-free) and cryopreservation, iPSC characterization, naïve reversion (feeder dependent and independent)
Gene editing- CRISPR/Cas9 guide RNA design, delivery and detection, traffic light reporter system, eGFP fusion-based reporter line generation
Differentiation- tri-lineage differentiation using EB and monolayer methods, neural, retinal and RPE differentiation, generation of retinal and cerebral organoids, derivation of mesodermal cell types like vascular progenitors, endothelial cells, pericytes, vascular smooth muscle cells, and osteogenic, chondrogenic and adipogenic differntiation
Molecular/CellBiology techniques-DNA, RNA, Protein isolation, RT-PCR, qPCR, western blot, flow cytometry, immunofluorescence, cryosectioning, Confocal imaging (LSM 710), MACS sorting, proliferation/senescence/apoptosis related assays
Microbiology- Bacterial transformation, plasmid cloning, isolation and transfection, lentiviral production and transduction in human cell lines
Animal experience- mice breeding and handling, teratoma injections and harvest, generation of mouse model of ischemic reperfusion injury (retina), and cell injection into vitreous body, harvesting mouse retina, primary culture from ear punch (mouse and rat), generation of irradiated mouse embryonic fibroblasts (MEFs) as feeders
Bioinformatics-In silico pathway analysis, microarray data analysis including DNA methylation by MeDIP CHIP, Ingenuity Pathway Analysis, miRNA expression analysis by nSolver and nCounter, primer design, sequence analysis
Howard University | | Washington , D.C, USAPh.D. in Anatomy2018
University of Mumbai | | Mumbai, Maharashtra, IndiaMS in Biotechnology2012
University of Pune | | Pune, Maharashtra, IndiaBS in Biotechnology2011
Kanherkar RR, Bhatia-Dey N, Makarev E and Csoka AB (2014) Cellular reprogramming for understanding and treating human disease. Front. Cell Dev. Biol. 2:67. doi:10.3389/fcell.2014.00067
Kanherkar RR, Bhatia-Dey N and Csoka AB (2014) Epigenetics across the human lifespan. Front. Cell Dev. Biol. 2:49. doi: 10.3389/fcell.2014.00049
Bhatia-Dey, N, Kanherkar, RR, Stair, SE, Makarev, EO, & Csoka, AB (2016) Cellular Senescence as the Causal Nexus of Aging. Frontiers in Genetics,7,13. doi.org/10.3389/fgene.2016.00013
Zhavoronkov A, Izumchenko E, Kanherkar RR, Teka M, Cantor C, Manaye K, Sidransky D, West MD, Makarev E, Csoka AB. (2016) Pro-fibrotic pathway activation in trabecular meshwork and lamina cribrosa is the main driving force of glaucoma. Cell Cycle (12) 1643-1652. doi:10.1080/15384101.2016.1170261.
Kanherkar RR, Stair SE, Bhatia-Dey N, Mills PJ, Chopra D, Csoka AB (2017) Epigenetic Mechanisms of Integrative Medicine. Evidence-Based Complementary and Alternative Medicine, vol. 2017, Article ID 4365429, 19 pages, 2017. doi:10.1155/2017/4365429
Makarev, E., Schubert, A.D., Kanherkar, R.R., London, N., Teka, M., Ozerov, I., Lezhnina, K., Bedi, A., Ravi, R., Mehra, R. and Hoque, M.O., (2017) In silico analysis of pathways activation landscape in oral squamous cell carcinoma and oral leukoplakia. Cell Death Discovery, 3, p.17022. doi: 10.1038/cddiscovery.2017.22
Kanherkar, R.R., Getachew, B., Ben-Sheetrit, J., Verma, S., Heinbockel, T., Tizabi, Y., Csoka, A.B. (2018) The effect of citalopram on genome-wide DNA methylation of human cells. International Journal of Genomics, vol 2018, Article ID 8929057, 12 pages, 2018. doi.org/10.1155/2018/8929057
National Institute of Health | Bethesda, MDGuest Lecturer08/2018 - 08/2018
Howard University | Washington , D.C.Graduate Teaching Assistant01/2017 - 12/2017
Howard University Research Day (Washington D.C. 2017) - Generation of induced pluripotent stem cell from Hutchinson-Gillford Progeria Syndrome patient fibroblasts using lentiviral, episomal and mRNA-based methods.
ISSCR annual meeting (Boston, MA 2017)- Generation of Induced pluripotent stem cells (iPSCs) from Hutchinson-Gilford Progeria Syndrome (HGPS) patient fibroblasts.
Advancing Computational Biology @ Howard University Symposium & Howard University Research Day (Washington D.C. 2016)- Pro-fibrotic pathway activation in trabecular meshwork and lamina cribrosa is the main driving force of glaucoma (Washington D.C. 2016).
Howard University Research Day (Washington D.C. 2015) - Epigenetic changes in human cells in response to chronic citalopram treatment.
Haffkine Institute Symposium (Mumbai, India 2012) - Assessment of neuronal oxidative stress in murine model of rabies
Fellowships and Awards
Merit -based Graduate scholarship- Howard University 2014-2018
Three year recipient of Lead Foundation scholarship with Academic Award for outstanding performance, for ranking number one in class during Bachelors Degree. (Pune, India, 2007-2010)
Peer-reviewed articles for (a) MedCrave's Online Journal of Stem cell Research and Therapeutics (JSRT), (b) German Society for Stem Cell Research's (GSZ)- Journal of Stem Cells & Regenerative Medicine (JSRM), (c) Nova Publishers' Journal of Stem Cells (JSC), (d) Life Sciences groups' Journal of Stem Cell Research & Therapy, and (e) Elsevier's Journal of Stem Cell Research (SCR).
Member of International Society for Stem Cell Research (ISSCR) Member (2017), Annual conference attendee (Boston, MA June 2017)
Member of The Association for Research in Vision and Ophthalmology (ARVO) Member (2017) and ARVO Annual conference attendee (Baltimore, MD May 2017)