THE PURIFICATION METHOD USING IODIXANOL (OPTIPREP)-BASED DENSITY GRADIENT SIGNIFICANTLY REDUCE CYTOKINE/CHEMOKINE PRODUCTION FROM HUMAN ISLET PREPARATIONS, LEADING TO PROLONG ∼ À-CELL SURVIVAL DURING CULTURE

2008 ◽  
Vol 86 (Supplement) ◽  
pp. 570
Author(s):  
A Mita ◽  
C Ricordi ◽  
A Miki ◽  
S Barker ◽  
A Khan ◽  
...  
Keyword(s):  
Cell Survival ◽  
Density Gradient ◽  
Human Islet ◽  
Purification Method ◽  
Chemokine Production ◽  
A Cell

scholarly journals A Maternal Smad Protein Regulates Early Embryonic Apoptosis in Xenopus laevis

2002 ◽  
Vol 22 (5) ◽  
pp. 1317-1328 ◽  
Author(s):  
Yuko Miyanaga ◽  
Ingrid Torregroza ◽  
Todd Evans
Keyword(s):  
Functional Analysis ◽  
Cell Survival ◽  
Caspase 3 ◽  
Embryonic Cell ◽  
Early Embryogenesis ◽  
Cell Cycles ◽  
Smad Proteins ◽  
Smad Protein ◽  
Smad Signaling Pathway ◽  
A Cell

ABSTRACT We identified cDNAs encoding the Xenopus Smad proteins most closely related to mammalian Smad8, and we present a functional analysis of this activity (also referred to recently as xSmad11). Misexpression experiments indicate that xSmad8(11) regulates pathways distinct from those regulated by the closely related xSmad1. Embryos that develop from eggs depleted of xSmad8(11) mRNA fail to gastrulate; instead, at the time of gastrulation, they initiate a widespread program of apoptosis, via a CPP32/caspase 3 pathway. Embryos that avoid this fate display gastrulation defects. Activation of apoptosis is rescued by expression of xSmad8(11) but not xSmad1. Our results demonstrate an embryonic requirement for Smad8(11) activity and show that a maternally derived Smad signaling pathway is required for gastrulation and for mediating a cell survival program during early embryogenesis. We suggest that xSmad8(11) functions as part of a maternally derived mechanism shown previously by others to monitor Xenopus early embryonic cell cycles.

Intersex-like (IXL) Is a Cell Survival Regulator in Pancreatic Cancer with 19q13 Amplification

2007 ◽  
Vol 67 (5) ◽  
pp. 1943-1949 ◽  
Author(s):  
Riina Kuuselo ◽  
Kimmo Savinainen ◽  
David O. Azorsa ◽  
Gargi D. Basu ◽  
Ritva Karhu ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cell Survival ◽  
A Cell

scholarly journals Comparison of Tissue Loading Before and After the Creation of a Continuous Density Gradient in Porcine Islet Purification

Cell Medicine ◽  
2018 ◽  
Vol 10 ◽  
pp. 215517901878134 ◽  
Author(s):  
Chika Miyagi-Shiohira ◽  
Yoshiki Nakashima ◽  
Nana Ebi ◽  
Eri Hamada ◽  
Yoshihito Tamaki ◽  
...  
Keyword(s):  
Density Gradient ◽  
Stimulation Index ◽  
High Yield ◽  
Pancreatic Islet Transplantation ◽  
Post Transplantation ◽  
Porcine Pancreas ◽  
Purification Method ◽  
Continuous Density ◽  
Porcine Islet ◽  
The Impact

The purification step is one of the most important and difficult procedures in islet isolation for pancreatic islet transplantation. We previously reported that a purification method using large plastic bottles effectively achieved a high yield of islets from the porcine pancreas. In this study, we evaluated the impact of the timing of tissue loading on porcine islet purification using large plastic bottles. One method involved loading digested tissue after creating a continuous density gradient (tissue after gradient [TAG]). The other method involved loading digested tissue before creating a continuous density gradient (tissue before gradient [TBG]). There were no significant differences between TAG and TBG in terms of the islet yield, rates of viability and purity, score, and in the stimulation index after purification. Furthermore, there were no marked differences in the attainability or suitability of post-transplantation normoglycemia. Our study shows the equivalency of these two methods of islet purification.

scholarly journals 2219

2017 ◽  
Vol 1 (S1) ◽  
pp. 59-59
Author(s):  
Shaheen Kurani ◽  
Nicolas Madigan ◽  
Karl Clark ◽  
Stephen Ekker ◽  
Nathan Staff ◽  
...  
Keyword(s):  
Cell Survival ◽  
Targeted Delivery ◽  
Transfection Efficiency ◽  
Current Treatment ◽  
Safe Harbor ◽  
Systemic Delivery ◽  
Transcription Activator ◽  
Field Methods ◽  
A Cell ◽  
The Central Nervous System

OBJECTIVES/SPECIFIC AIMS: The current treatment for amyotrophic lateral sclerosis (ALS) includes systemic delivery of neurotrophic factors (NTFs). Although this approach may seem theoretically sound, NTF efficacy within the central nervous system (CNS) is largely limited by the blood-brain barrier. Thus, a cell-based approach, which allows for targeted delivery of molecular therapies locally from the CNS, could lead to a paradigm shift in the field. METHODS/STUDY POPULATION: The Windebank and Staff group at Mayo Clinic completed a Phase I dose-escalation safety trial of autologous, adipose-derived mesenchymal stem cells (adMSCs) in an effort to move toward personalized medical treatment of ALS. The adMSCs were injected into the intrathecal space by lumbar puncture in 27 patients and the results showed an excellent safety profile across a range of doses. The team is moving forward with this idea by using gene-editing technology to develop clinical-grade, genetically modified autologous MSCs. The patient-derived adMSCs are modified at defined “safe-harbor” regions of the human genome through transcription activator-like effector nuclease (TALEN) technology. RESULTS/ANTICIPATED RESULTS: Our results show that electroporating adMSCs with plasmid DNA leads to efficient GFP or TALEN transgene expression, but yields low cell survival and a low rate of genetic modification. DISCUSSION/SIGNIFICANCE OF IMPACT: It can be concluded that: (1) TALEN technology may be used to target safe harbor loci for gene integration to produce therapeutic adMSC for ALS. (2) Primary barriers to adMSC modification are inefficient TALEN and donor template uptake, low cutting efficiency, and poor cell survival after electroporation. Future directions include optimizing the protocol to obtain 48 base pairs in the homology arms and increasing transfection efficiency.

scholarly journals Identification of Proteins Associating with Glycosylphosphatidylinositol- Anchored T-Cadherin on the Surface of Vascular Endothelial Cells: Role for Grp78/BiP in T-Cadherin-Dependent Cell Survival

2008 ◽  
Vol 28 (12) ◽  
pp. 4004-4017 ◽  
Author(s):  
Maria Philippova ◽  
Danila Ivanov ◽  
Manjunath B. Joshi ◽  
Emmanouil Kyriakakis ◽  
Katharina Rupp ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Surface ◽  
Cell Survival ◽  
High Performance ◽  
Vascular Endothelial Cells ◽  
Surface Lipid ◽  
Gaba A ◽  
Dependent Cell ◽  
A Cell ◽  
Cell Surface Signaling

ABSTRACT There is scant knowledge regarding how cell surface lipid-anchored T-cadherin (T-cad) transmits signals through the plasma membrane to its intracellular targets. This study aimed to identify membrane proteins colocalizing with atypical glycosylphosphatidylinositol (GPI)-anchored T-cad on the surface of endothelial cells and to evaluate their role as signaling adaptors for T-cad. Application of coimmunoprecipitation from endothelial cells expressing c-myc-tagged T-cad and high-performance liquid chromatography revealed putative association of T-cad with the following proteins: glucose-related protein GRP78, GABA-A receptor α1 subunit, integrin β3, and two hypothetical proteins, LOC124245 and FLJ32070. Association of Grp78 and integrin β3 with T-cad on the cell surface was confirmed by surface biotinylation and reciprocal immunoprecipitation and by confocal microscopy. Use of anti-Grp78 blocking antibodies, Grp78 small interfering RNA, and coexpression of constitutively active Akt demonstrated an essential role for surface Grp78 in T-cad-dependent survival signal transduction via Akt in endothelial cells. The findings herein are relevant in the context of both the identification of transmembrane signaling partners for GPI-anchored T-cad as well as the demonstration of a novel mechanism whereby Grp78 can influence endothelial cell survival as a cell surface signaling receptor rather than an intracellular chaperone.

scholarly journals Immunolocalization of the Bcl-2 protein within hematopoietic neoplasms

Blood ◽  
1991 ◽  
Vol 78 (4) ◽  
pp. 1062-1068 ◽  
Author(s):  
M Zutter ◽  
D Hockenbery ◽  
GA Silverman ◽  
SJ Korsmeyer
Keyword(s):  
Cell Survival ◽  
Large Cell ◽  
Hematologic Malignancies ◽  
Lymphocytic Leukemia ◽  
Myelogenous Leukemia ◽  
Protein Levels ◽  
Stage Of Development ◽  
T Cell Lymphomas ◽  
A Cell ◽  
Reactive Hyperplasia

Abstract The Bcl-2 proto-oncogene was discovered at the t(14;18) breakpoint found in most follicular B-cell lymphomas and some diffuse large-cell lymphomas. Bcl-2 is unique among proto-oncogenes, being localized to mitochondria and extending cell survival by blocking programmed cell death. We examined Bcl-2 protein expression in 82 hematologic malignancies and reactive lymphoid processes. All lymphomas with Bcl-2 rearrangement demonstrated high levels of Bcl-2 protein. However, most follicular and diffuse lymphomas without Bcl-2 rearrangement also displayed intense Bcl-2 staining. In these cases, mechanisms other than classic translocation may be deregulation Bcl-2. The pattern of Bcl-2 staining in follicular lymphoma is the inverse of the pattern in reactive hyperplasia, confirming a role for Bcl-2 immunolocalization in routine diagnosis. Small lymphocytic malignancies, including small lymphocytic lymphoma, mantle zone lymphoma, and chronic lymphocytic leukemia, expressed intermediate levels of Bcl-2. Bcl-2 protein varied in plasma cell dyscrasias. Bcl-2 protein levels in T-cell lymphomas reflected their corresponding stage of development. No substantial Bcl- 2 was present in the Reed-Sternberg cells of nodular sclerosing Hodgkin's disease. Chronic myelogenous leukemia was strongly positive for Bcl-2, consistent with the presence of Bcl-2 in normal myeloid progenitors. Immunohistochemistry identified an expanded spectrum of hematopoietic neoplasms in which Bcl-2 may provide a cell survival advantage.

scholarly journals A Clinically Applicable Positive Allosteric Modulator of GABA Receptors Promotes Human β-Cell Replication and Survival as well as GABA’s Ability to Inhibit Inflammatory T Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-7 ◽  
Author(s):  
Jide Tian ◽  
Hoa Dang ◽  
Nataliya Karashchuk ◽  
Irvin Xu ◽  
Daniel L. Kaufman
Keyword(s):  
T Cell ◽  
Cell Survival ◽  
Islet Cell ◽  
Cell Mass ◽  
T Cell Responses ◽  
Human Islet ◽  
Cell Replication ◽  
Cell Responses ◽  
Autoreactive T Cell

A major goal of T1D research is to develop new approaches to increase β-cell mass and control autoreactive T cell responses. GABAA-receptors (GABAA-Rs) are promising drug targets in both those regards due to their abilities to promote β-cell replication and survival, as well as inhibit autoreactive T cell responses. We previously showed that positive allosteric modulators (PAMs) of GABAA-Rs could promote rat β-cell line INS-1 and human islet cell replication in vitro. Here, we assessed whether treatment with alprazolam, a widely prescribed GABAA-R PAM, could promote β-cell survival and replication in human islets after implantation into NOD/scid mice. We observed that alprazolam treatment significantly reduced human islet cell apoptosis following transplantation and increased β-cell replication in the xenografts. Evidently, the GABAA-R PAM works in conjunction with GABA secreted from β-cells to increase β-cell survival and replication. Treatment with both the PAM and GABA further enhanced human β-cell replication. Alprazolam also augmented the ability of suboptimal doses of GABA to inhibit antigen-specific T cell responses in vitro. Thus, combined GABAA-R agonist and PAM treatment may help control inflammatory immune responses using reduced drug dosages. Together, these findings suggest that GABAA-R PAMs represent a promising drug class for safely modulating islet cells toward beneficial outcomes to help prevent or reverse T1D and, together with a GABAA-R agonist, may have broader applications for ameliorating other disorders in which inflammation contributes to the disease process.

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