| 1. |
|
|
| 2. |
|
|
| 3. |
- Yao, M, et al.
(author)
-
Cibinetide Protects Isolated Human Islets in a Stressful Environment and Improves Engraftment in the Perspective of Intra Portal Islet Transplantation
- 2021
-
In: Cell transplantation. - : SAGE Publications. - 1555-3892 .- 0963-6897. ; 30, s. 9636897211039739-
-
Journal article (peer-reviewed)abstract
- During intra-portal pancreatic islet transplantation (PITx), innate immune reactions such as the instant blood mediated inflammatory reaction (IBMIR) cause an immediate loss of islets. The non-hematopoietic erythropoietin analogue cibinetide has previously shown islet-protective effects in mouse PITx. Herein, we aimed to confirm cibinetide’s efficacy on human islets, and to characterize its effect on IBMIR. We cultured human islets with pro-inflammatory cytokines for 18 hours with or without cibinetide. ATP content and caspase 3/7 activity were measured. Dynamic glucose perfusion assay was used to evaluate islet function. To evaluate cibinetides effect on IBMIR, human islets were incubated in heparinized polyvinyl chloride tubing system with ABO compatible blood and rotated for 60 minutes to mimic the portal vein system. Moreover, human islets were transplanted into athymic mice livers via the portal vein with or without perioperative cibinetide treatment. The mice were sacrificed six days following transplantation and the livers were analyzed for human insulin and serum for human C-peptide levels. Histological examination of recipient livers to evaluate islet graft infiltration by CD11b+ cells was performed. Our results show that cibinetide maintained human islet ATP levels and reduced the caspase 3/7 activity during culture with pro-inflammatory cytokines and improved their insulin secreting capacity. In the PVC loop system, administration of cibinetide reduced the IBMIR-induced platelet consumption. In human islet to athymic mice PITx, cibinetide treatment showed an increased amount of human insulin in the livers and higher serum human C-peptide, while histological examination of the livers showed reduced infiltration of pro-inflammatory CD11b+ cells around islets grafts compared to the controls. In summary, Cibinetide protected isolated human islets in a pro-inflammatory milieu and reduced IBMIR related platelet consumption. It improved engraftment of human islets in athymic mice. The study confirms that cibinetide is a promising agent to be used in clinical PITx.
|
|
| 4. |
- Akhmanova, M, et al.
(author)
-
Physical, Spatial, and Molecular Aspects of Extracellular Matrix of In Vivo Niches and Artificial Scaffolds Relevant to Stem Cells Research
- 2015
-
In: Stem cells international. - : Hindawi Limited. - 1687-966X .- 1687-9678. ; 2015, s. 167025-
-
Journal article (peer-reviewed)abstract
- Extracellular matrix can influence stem cell choices, such as self-renewal, quiescence, migration, proliferation, phenotype maintenance, differentiation, or apoptosis. Three aspects of extracellular matrix were extensively studied during the last decade: physical properties, spatial presentation of adhesive epitopes, and molecular complexity. Over 15 different parameters have been shown to influence stem cell choices. Physical aspects include stiffness (or elasticity), viscoelasticity, pore size, porosity, amplitude and frequency of static and dynamic deformations applied to the matrix. Spatial aspects include scaffold dimensionality (2D or 3D) and thickness; cell polarity; area, shape, and microscale topography of cell adhesion surface; epitope concentration, epitope clustering characteristics (number of epitopes per cluster, spacing between epitopes within cluster, spacing between separate clusters, cluster patterns, and level of disorder in epitope arrangement), and nanotopography. Biochemical characteristics of natural extracellular matrix molecules regard diversity and structural complexity of matrix molecules, affinity and specificity of epitope interaction with cell receptors, role of non-affinity domains, complexity of supramolecular organization, and co-signaling by growth factors or matrix epitopes. Synergy between several matrix aspects enables stem cells to retain their function in vivo and may be a key to generation of long-term, robust, and effective in vitro stem cell culture systems.
|
|
| 5. |
- Domogatskaya, A, et al.
(author)
-
Functional diversity of laminins
- 2012
-
In: Annual review of cell and developmental biology. - : Annual Reviews. - 1530-8995 .- 1081-0706. ; 28, s. 523-553
-
Journal article (peer-reviewed)abstract
- Laminins are a large family of conserved, multidomain trimeric basement membrane proteins that contribute to the structure of extracellular matrix and influence the behavior of associated cells, such as adhesion, differentiation, migration, phenotype stability, and resistance to anoikis. In lower organisms such as Hydra there is only one isoform of laminin, but higher organisms have at least 16 trimeric isoforms with varying degrees of cell/tissue specificity. In vitro protein and cell culture studies, gene manipulation in animals, and laminin gene mutations in human diseases have provided insight into the specific functions of some laminins, but the biological roles of many isoforms are still largely unexplored, mainly owing to difficulties in isolating them in pure form from tissues or cells. In this review, we elucidate the evolution of laminins, describe their molecular complexity, and explore the current knowledge of their diversity and functional aspects, including laminin-mediated signaling via membrane receptors, in vitro cell biology, and involvement in various tissues gained from animal model and human disease studies. The potential use of laminins in cell biology research and biotechnology is discussed.
|
|
| 6. |
|
|
| 7. |
|
|
| 8. |
- Ojala, JRM, et al.
(author)
-
A novel scavenger receptor 5-based antibiotic-independent selection method for generation of stable recombinant protein-producing mammalian cell lines especially suitable for proteins affecting cell adhesion
- 2012
-
In: BioTechniques. - : Future Science Ltd. - 1940-9818 .- 0736-6205. ; 53:4, s. 221-
-
Journal article (peer-reviewed)abstract
- The establishment of stable recombinant protein-producing mammalian cell lines is an expensive, time-consuming, tedious procedure. In some cases, expressed recombinant proteins have adverse effects on host cell function, including cell adhesion. Based on the adhesive properties of SCARA5, a scavenger receptor (SR) of the class A SR family, we developed a method for selection of stable recombinant protein-producing cell clones that relies on an internal ribosome entry site (IRES) vector where the protein of interest is expressed in the same messenger RNA as SCARA5, resulting in improved adhesion and increased cell viability of recombinant protein-producing cells in serum-free media. This method does not depend on antibiotics, complicated selective cell culture media or equipment, and thus offers the advantages of being inexpensive, environmentally friendly, and simple.
|
|
| 9. |
|
|
| 10. |
- Rodin, S, et al.
(author)
-
Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment
- 2014
-
In: Nature communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 5, s. 3195-
-
Journal article (peer-reviewed)abstract
- Lack of robust methods for establishment and expansion of pluripotent human embryonic stem (hES) cells still hampers development of cell therapy. Laminins (LN) are a family of highly cell-type specific basement membrane proteins important for cell adhesion, differentiation, migration and phenotype stability. Here we produce and isolate a human recombinant LN-521 isoform and develop a cell culture matrix containing LN-521 and E-cadherin, which both localize to stem cell niches in vivo. This matrix allows clonal derivation, clonal survival and long-term self-renewal of hES cells under completely chemically defined and xeno-free conditions without ROCK inhibitors. Neither LN-521 nor E-cadherin alone enable clonal survival of hES cells. The LN-521/E-cadherin matrix allows hES cell line derivation from blastocyst inner cell mass and single blastomere cells without a need to destroy the embryo. This method can facilitate the generation of hES cell lines for development of different cell types for regenerative medicine purposes.
|
|
