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International Journal of Translational Science


Pranela Rameshwar, Rutgers University, USA
Paolo Di Nardo, Rome Tor Vergata, Italy

ISSN: 2246-8765 (Online Version)
Vol: 2016   Issue: 1

Published In:   January 2016

Publication Frequency: Continuous Article Publication

Articles in 2020

Search Available Volume and Issue for International Journal of Translational Science

Characterization of Mesenchymal Stem Cells from Human Cortical Bone

Joseph S. Fernandez-Moure1, Bruna Corradetti2, Trevor Janecek1, Jeffrey Van Eps1, Matthew Burn1, Bradley K.Weiner1, Pranela Rameshwar3 and Ennio Tasciotti2

1The Methodist Hospital Research Institute, Department of Surgery, Houston Methodist Hospital, Houston, Texas, USA
2Houston Methodist Research Institue, Houston, Texas, USA
3Rutgers New Jersey Medical School, Newark, New Jersey, USA

Abstract: [+]    |    Download File [ 2875KB ]    |   Read Article Online

Background Context: Mesenchymal stem cells (MSC) are being used for spine and orthopaedic surgical and research applications. Bone marrow and fat are the most commonly used sources of these cells.

Purpose: To describe a new technique allowing the isolation and expansion of human MSC from cortical bone.

Study Design: MSC from human cortical bone (vertebral lamina) were isolated, expanded, and verified in vitro.

Methods: Human MSC were isolated from laminar bone obtained during surgery (decompression/laminectomy). They were then cultured and assessed using fluorescence-activated cell sorting techniques forMSCmarkers, colonyforming unit assays, and multilineage differentiation.

Results: Isolated and cultured cells demonstrated MSC markers and trilineage differentiation confirming their stemness.

Conclusion: Anovel method for the isolation of MSC from cortical bone has been described. These cells have significant current and future application in spine and orthopaedic surgery; and both the source of the cells and particular characteristics of the cortical bone derived MSC have advantages over currently used MSC obtained from other sources.

Levels of Stromal Derived Factor-1 (SDF-1) and Brain Derived Neurotropic Factor (BDNF) and Very Small Embryonic-Like Cells (VSEL) in Ischemic Stroke Patients

Bayu Winata Putera1,2, Cynthia R. Sartika2, Andi Wijaya1,2,2 Irawan Yusuf1 and Aw Tar-Choon1,4

1Postgraduate Program in Clinical Biochemistry, Hasanuddin University, Indonesia
2Prodia Stemcell Indonesia
3Prodia Clinical Laboratory
4National University of Singapore

Abstract: [+]    |    Download File [ 1230KB ]    |   Read Article Online

Abstract: Ischemic stroke remains a major health problem associated with high mortality and severe morbidity. The challenge of treatment is to understand the process leading to endogenous neurorepair mechanism to ischemic stroke. This study tested the hypothesis that VSEL, SDF-1 and BDNF have important roles in the process endogenous neurorepair in response to ischemic stroke. Studies indicated an increase in SDF-1 and VSEL within one week of stroke. BDNF levels tapered after day 15. Together the studies indicated that BDNF levels were highest when measured within 7 days of stroke onset and decreased thereafter. SDF-1 and VSEL were highest at between 7 and 15 days of stroke onset. The findings indicated that SDF-1 could be key for VSEL to be mobilized as a natural repair process whereas BDNF might be the correlative response to prevent cell death.

Keywords: SDF-1, BDNF, VSEL, Ischaemic stroke.

Cancer Vaccines: Bench to Bedside

Neha Tuli†,1, Rachana Maniyar†,1, Robert Bednarczyk1, Ghada Ben Rahoma1, Sarnath Singh1, Jan Geliebter1, Abraham Mittelma1, Marc Wallack1,2, Debabrata Banerjee3 and Raj K. Tiwari1

1Departmentof Microbiology and Immunology, New York Medical College, Valhalla, NY 10595, USA
2Department of Surgery, Metropolitan Hospital Centre 1901 1st Avenue, New York, NY 10029, USA
3Department of Pharmacology, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA
These authors contributed equally to the preparation of this manuscript

Abstract: [+]    |    Download File [ 4102KB ]    |   Read Article Online

Abstract: The immune system has immense potential in cancer therapy as it is individualized, precision driven and robust, however, it is associated with challenges of its own that include immune evasion, development of tolerance and a sustained tumor rejection response. Recent FDA approval of several checkpoint inhibitors, anti-CTLA4, anti-PD-1, has re-invigorated cancer immunology by demonstrating that tolerance to cancer can be broken to induce a sustained immune response in patients. Active immunization with multivalent tumor associated antigens (TAA), however, is still a challenge. We have developed two specific distinct methods to generate multivalent antigens capable of tumor regression in prostate cancer and melanoma. In prostate cancer,we have generated specific multivalent peptide mimetics using phage display synthetic peptide libraries capable of metastatic tumor regression in an animal model. In melanoma, we have used a vaccinia virus based antigen retrieval technology to generate a multivalent antigenic vaccine. The antigenic repertoire is well defined. A protocol for the melanoma vaccine is FDA approved for clinicaltrials. We envision defining the humoral and cellular immune response to combine our active vaccine strategy with other treatment modalities including approved checkpoint inhibitors anti-CTLA4 and anti-PD-1. We believe our vaccine candidates are a new generation of immune-therapeutics that can prolong cancer free survival and prevent secondary recurrences. Our studies have challenged the existing paradigms to re-define cancer immunotherapy that bridges the gap between humoral and cellular immunity by combining active immune response with negative checkpoint inhibitors thus activating pre-existing dormant immune response.

Keywords: cancer vaccines, peptide epitopes, vaccinia virus – antigen retrieval.

Keratinocyte Induced Differentiation of Mesenchymal Stem Cells into Dermal Myofibroblasts: A Role in EffectiveWound Healing

Pravin J. Mishra1, Prasun J. Mishra2 and Debabrata Banerjee3

1Intermountain Precision Genomics, Intermountain Healthcare, Dixie Regional Medical Center 292 South 1470 East, Suite 201 & 301, St. George, UT 84770, USA
2Department of Biochemical and Cellular Pharmacology, Genentech, 1, DNA Way, South San Francisco, California 94080, USA
3Department of Pharmacology, Robert Wood Johnson Medical School, Graduate School of Biomedical Sciences, New Brunswick-Piscataway, Rutgers University, 675 Hoes Lane West, Piscataway, NJ 08854. USA

Abstract: [+]    |    Download File [ 3349KB ]    |   Read Article Online

Abstract: We have previously demonstrated that human mesenchymal stem cells (hMSCs) migrate toward human keratinocytes as well as toward conditioned medium from cultured human keratinocytes (KCM) indicating that the hMSCs respond to signals from keratinocytes [1]. Using fluorescently labeled cells we now show that in vitro hMSCs appear to surround keratinocytes, and this organization is recapitulated in vivo. Incubation of hMSCs with KCM induced dermal myofibroblast like differentiation characterized by expression of cytoskeletal markers and increased expression of cytokines including SDF-1, IL-8, IL-6 and CXCL5. Interaction of keratinocytes with hMSCs appears to be important in the wound healing process. Therapeutic efficacy of hMSCs in wound healing was examined in two animal models representing normal and chronic wound healing. Accelerated wound healing was observed when hMSCs and KCM exposed hMSCs (KCMSCs) were injected near wound site in nude and NOD/SCID mice. Long term follow up of wound healing revealed that in the hMSC treated wounds there was little evidence of residual scarring. These dermal myofibroblast like hMSCs add to the wound healing process. Together, the keratinocyte and hMSCs morphed dermal myofibroblast like cells as well as the factors secreted by these cells support wound healing with minimal scarring. The ability of hMSCs to support wound healing process represents another striking example of the importance of keratinocyte and hMSCs interplay in the wound microenvironment resulting in effective wound healing with minimal scarring.

Keywords: Mesenchymal stem cells, keratinocyte conditioned medium, dermal myofibroblast, cytokine secretion, wound healing, scarring, animal models.

Immunoassays for Affinity, Avidity and Competition: Methods to Evaluate Monoclonal (and Polyclonal) Antibodies Useful Assays in Development of New Antibodies, Better Understanding of Commercial Antibodies: Their Usefulness in Specific Aims of Research [SOP]

Anita Lewis Antes

New Jersey Medical School/Rutgers University/Newark NJ, USA

Abstract: [+]    |    Download File [ 122KB ]    |   Read Article Online

Abstract: Enzyme-linked immunosorbent assay (ELISA) remains the most common method to identify clones of cells during the development of monoclonal antibodies. This technique is convenient for the rapid screening of large numbers of clones, subsequent to the fusion of splenocytes of immunized mice or rabbits with immortal myeloma cells. In general, when screening for the production of the desired antibody, an antibody to the target protein or an antibody from another host is generally unavailable. Thus, the standard test measures reactivity of attachment passively to the target antigen, and reporting of that attachment using a secondary antibody reporter (sandwich assay). Slightly better is the ELISA, which reacts the monoclonal within the cell culture (supernatant) with the antigen and then applies the mix to plates coated with an antibody against the target, but from another host species. In this way, the ability of the test clones to capture the target may be examined. This method is required when preparing antibodies for immunoprecipitation.

The sandwich assay may leave the investigator with as many as 100 or more prospective clones to further characterize. To identify clones for further investigation of the target antigen, cell supernatants may be too dilute. Thus, the expansion of this enormous number of clones for subsequent purification of the antibodies would be a daunting task. Here we offer EIA-based assays which simplify the process of excluding the majority of clones which will not serve the researcher’s requirements. All the assays can be done from a single stock of each clone of as little as 2 ml volume.

Keywords: ELISA (enzyme linked immune absorbant assay), specificity, sensitivity, affinity, monoclonal antibody, hybridoma, epitope.

River Publishers: International Journal of Translational Science