In the human body, vascular injuries that are caused by trauma, vessel lumen stenosis, and occlusions are often irreversible and can lead to sequelae formation as the vessels cannot reproduce fast enough. To solve this problem, the blood flow must be returned to the region as fast as possible. The adipose tissue contains progenitor cells with angiogenic potential and can be used to resolve the issue. In the present study, mesenchymal stem cells (MSCs) derived from rat adipose tissue, vascular endothelial growth factor (VEGF), and their mixture were applied on the dorsum of a rat, which was traumatized and its contribution to vascular regeneration was reviewed. No application was made to the control group. The results showed that the percentage of necrotic area was significantly lower in the MSC group than that of all the other groups. When the VEGF group was compared to the VEGF + MSCs, the percentage of necrotic area was observed to be similiar. However, VEGF showed effects only when a large quantites of VEGF was applied to the flap area. VEGF could not fully respond to the needs, whereas MSCs can produce VEGF according to the needs of tissue. This makes them superior to stem cells.
A new polyethylene glycol (PEG) based microcarrier was designed and examined by the attachment and growth of mouse fibroblast cells. In the design of microcarrier, a PEG-based macromonomer, polyethyleneglycol methacrylate (PEGMA), was selected as the main component of hydrogel beads since PEG is known as a nontoxic and biocompatible material. A relatively new cationic comonomer, N-[3-(dimethylamino)propyl]methacrylamide (DMAPM), with higher ionization ability with respect to the similar comonomers was used for providing cationic charge to the hydrogel structure. In the first part, a suspension copolymerization method was developed for the production of cationically charged hydrogel beads as a potential microcarrier for cell culturing. The suspension copolymerization by using ethylene dimethacrylate (EDM) as cross-linking agent and cyclohexanol as the diluent provided spherical, polydisperse poly(PEGMA-DMAPM-EDM) hydrogel beads with an average size of 121 microm. The hydrogel beads exhibited a pH-dependent swelling behavior. The L929 mouse fibroblast cells were cultured on poly(PEGMA-DMAPM-EDM) hydrogel beads with an initial concentration of 200,000 cells/mL. The cells were incubated in Dulbecco's modified Eagle's medium during 5 days and the cell proliferation was investigated at every 24 h. An effective cell attachment and growth up to 3.5 x 10(6) cells/mL were observed with the poly(PEGMA-DMAPM-EDM) hydrogel beads. The results indicated that the proposed microcarrier was a significant alternative to the hydrogel beads obtained by the copolymerization of 2-hydroxyethyl methacrylate and 2-dimethylaminoethylmethacrylate commonly used in microcarrier-facilitated cell culturing studies.
BackgroundFamilial hemophagocytic lymphohistiocytosis 2 (FHL2) is the most common familial type of hemophagocytic lymphohistiocytosis with immune dysregulation. FHL2 patients have mutations in the perforin gene which cause overactivation and proliferation of cytotoxic T lymphocytes and natural killer cells. Perforin is the key component of the cytolytic granule response function of cytotoxic T lymphocytes and natural killer cells. Perforin dysfunction causes a cytotoxic immune deficiency with a clinical outcome of uncontrolled and continuous immune stimulation response. This excessive stimulation leads to continuous systemic inflammation and, ultimately, multiorgan failure. Radical therapy is hematopoietic stem cell transplantation which is limited by the availability of a donor. Exacerbations of inflammatory attacks require a palliative immunosuppressive regimen. There is a need for an alternative or adjuvant therapy to maintain these patients when immunosuppression is ineffective or a donor is not available. Beneficial actions of mesenchymal stem cells (MSCs) have been shown in autoimmune diseases in clinical trials and are attributed to their immune-modulatory properties. This study aimed to assess the immune-modulatory effect of MSCs in an in-vitro model of FHL2.MethodsWe generated a targeted mutation in the perforin gene of NK92 cells to create an in-vitro FLH2 model using Crispr/Cas technology. A coculture setup was employed to assess the immunomodulatory efficacy of MSCs.ResultsEngineered NK92 clones did not show PRF1 mRNA expression and failed to secrete perforin upon phorbol myristate acetate–ionomycin stimulation, providing evidence for a valid FHL2 model. Coculture media of the engineered cells were investigated for the abundance of several cytokines. Coculture with MSCs revealed a reduction in major proinflammatory cytokines and an induction in anti-inflammatory and immunomodulatory cytokines compared to the parental NK92 cells.ConclusionsThis study shows the ameliorating effect of MSCs as an adjuvant immune modulator toward the therapy of FHL2 patients. MSCs are supportive therapy candidates for FHL2 patients under circumstances where prolonged immunosuppression is required to gain time before allogeneic hematopoietic stem cell transplantation.Electronic supplementary materialThe online version of this article (10.1186/s13287-018-0941-y) contains supplementary material, which is available to authorized users.
In this study, attachment and growth of Baby Hamster Kidney (BHK) cells on ethylene diamine (EDA)-plasma-treated poly(L-lactide/epsilon-caprolactone) biodegradable copolymer films were investigated. The co-polymer (Mw: 58000; Mn: 35000 and PI 1.60) was synthesised by ring-opening polymerization of the respective dimers with using stannous octoate as the catalyst. The final ratio of L-lactide to epsilon-caprolactone obtained by 1H-NMR was 87:13. The co-polymer films were treated with the EDA-plasma in a glow-discharge apparatus. The BHK-30 cell line was cultured on plain and EDA-plasma-treated films and their pre-wetted forms (with ethanol and/or cell culture medium before use). Cell attachment and growth were followed. Alkaline phosphatase (ALP) activity and glucose uptake in cell culture medium were also investigated. There was no attachment in the first 12 h. Glow-discharge treatment increased significantly the attachment and growth. Pre-wetting with ethanol and cell culture medium was also increase significantly both the attachment and growth.
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