Isolation and Expansion of Mesenchymal Stem Cells from Human Conjunctival Tissue

Nadri, Samad; Yazdani, Shahin

doi:10.1002/9780470151808.sc01f14s33

Cited by 3 publications

(3 citation statements)

References 24 publications

(27 reference statements)

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“…In line with previous studies, our MTT assay results illustrated that viability of CJMSCs cultured on fibrin gel significantly enhanced relative to cells cultured on TCP surface. This might be due to the polymerization processes of the fibrin gel that can facilitate oxygen and nutrients diffusion for the transplanted cells [12,34,39,40]. In this study, we found that culture of CJMSCs in fibrin gel leads to promotion of CJMSCs differentiation into photoreceptor-like cells.…”

Section: Discussionmentioning

confidence: 63%

“…Also, the qRT-PCR results showed expression rhodopsin, CRX and recoverin but at higher rates in the 3 D group. The results of immunocytochemistry and real-time PCR represented the higher level of differentiation to photoreceptor-like cells in cultured cells on Fibrin gel scaffold relative to those on a TPC surface (2 D control group) [33,39,40].…”

Section: Discussionmentioning

confidence: 99%

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Fibrin gel as a scaffold for photoreceptor cells differentiation from conjunctiva mesenchymal stem cells in retina tissue engineering

Soleimannejad

Ebrahimi‐Barough

Soleimani

et al. 2017

Artificial Cells, Nanomedicine, and Biotechnology

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Stem cell-based therapies are attraction approaches for regenerative medicine for treating retinal diseases. One of the limitations in cell therapy is cell death following post-injection whit preventing functional integration with retinal tissue. Fibrin gel, a bio-polymeric material with excellent biocompatibility, provides numerous advantages as a tissue engineering scaffold and a stem cell carrier. Therefore, current research is focusing on developing fibrin hydrogel scaffolds to protect stem cells during delivery and to stimulate endogenous regeneration through interactions of transplanted stem cells and retinal tissue. In this study fibrin gel was used as hydrogel scaffold for immobilization of cells. The structural characteristics of fibrin gel scaffold were examined with SEM. Rheological properties of fibrin gel were measured by rheometer and biodegradation rate of fibrin were assayed for 2 weeks. After isolation of stem cells CJMSCs, the cells were differentiated into photoreceptor-like cells by exposing with taurin for 14 days in tissue culture plate (TCP group) and fibrin hydrogel (3 D group). The attachment of cells was analyzed with SEM and MTT. The expression of rhodopsin, PKC, CRX, recoverin, peripherin, nestin and RPE65 as photoreceptor-like cell markers was evaluated by immunocytochemistry and quantitative real-time PCR (RT-PCR) in TCP and 3 D groups. The results of SEM analysis showed CJMSCs were well attached in fibrin gels and there were good integrity between cells and scaffold. The elastic modulus and constant degradation of the gel contributes to the growth and proliferation of cells. There was no toxicity effect of fibrin hydrogel on cells and the viability of cultured cells was higher in 3 D fibrin gels in comparison with TCP groups. After 2 weeks, the expression of rhodopsin, PKC, CRX, peripherin, recoverin, nestin and RPE65 as special markers of photoreceptor cells were detected by Real time PCR and immunofluorescence that these expressions in 3 D groups were higher than TCP groups. In conclusion, our findings showed that application of readily available sources of adult stem cells like human conjunctiva stem cells encapsulated in fibrin gel could be interesting strategy to enhance photoreceptor progenitor cell numbers for repair and regeneration of retina disease such as photoreceptor injury.

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Section: Discussionmentioning

confidence: 63%

Section: Discussionmentioning

confidence: 99%

Fibrin gel as a scaffold for photoreceptor cells differentiation from conjunctiva mesenchymal stem cells in retina tissue engineering

Soleimannejad

Ebrahimi‐Barough

Soleimani

et al. 2017

Artificial Cells, Nanomedicine, and Biotechnology

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“…CJMSCs were isolated according to a protocol modified by Nadri et al 17,18 In brief, 2-3 mm 2 of conjunctiva biopsy ([eye globes] was obtained from the Central Eye Bank of I. R. Iran) was treated in supplemented hormonal epithelial medium (SHEM) containing 50 mg/mL dispase II (Sigma Chemical Co.) and 100 mM sorbitol. Under a stereomicroscope, epithelial sheets were separated and the isolated stromal tissue segments were cultured in DMEM/F-12 (1:1) (GIBCO-BRL) supplemented with 10% knockout serum (GIBCO-BRL), 4 ng/mL basic-FGF (Peprotech), 5 mg/mL insulin (Sigma Chemical Co.), and 10 ng/mL human LIF (Chemicon, Temecula, CA) and were incubated at 378C with 5%CO 2 in a humidified chamber.…”

Section: Isolation and Differentiation Into Mesenchymal Lineage Cellsmentioning

confidence: 99%

Effect of parameters on the quality of core‐shell fibrous scaffold for retinal differentiation of conjunctiva mesenchymal stem cells

Nadri

Nasehi

Barati

2016

J Biomedical Materials Res

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This article describes the coaxial electrospinning to generate core-shell fibers from polycaprolactone (PCL) and polyethylene glycol (PEG) for differentiation of conjunctiva mesenchymal stem cells (CJMSCs) into photoreceptor-like cells by delivery of taurine. Also, the effects of many parameters such as polymer concentration, nozzle collector distance, applied voltage, and outer solution flow rate on creation of the core-shell structure were examined. The morphology and structure of fibers were characterized using scanning, transmission electron microscopy, and fourier transform infrared spectroscopy and then retinal differentiation was examined by quantitative real time PCR (qPCR). Significant variations between 25% and 35% PEG concentration groups for fiber diameter were documented. As faster flowing rates from the outer nozzle (PCL fluid) were applied, the creation possibility of fibrous scaffold was increased. The lowest diameter and the best quality alignment of core-shell fibrous scaffold were achieved in 22 Kv and 24 Kv. As rising distancing were applied, the fibrous diameter increased and spraying was observed. qPCR analysis demonstrated the differentiation of CJMSCs to photoreceptor like cells on PEG/PCL scaffolds. According to the result, we have proved successful in the creation of core/shell fibrous scaffold of PEG/PCL by coaxial electrospinning for retinal tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 189-197, 2017.

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Differentiation of conjunctiva mesenchymal stem cells into secreting islet beta cells on plasma treated electrospun nanofibrous scaffold

Nadri

Barati

Mostafavi

et al. 2017

Artificial Cells, Nanomedicine, and Biotechnology

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Transplantation of stem cells using biocompatible nanofibrous scaffolds is a promising therapeutic method for treating Diabetic Mellitus. The aim of this study was to derive insulin-producing cells (IPCs) from conjunctiva-derived mesenchymal stem cell (CJMSCs) and to compare the functionality of differentiated IPCs in a three-dimensional (3D) culture with 2D. Furthermore, the effects of hydrophobicity of scaffold on IPCs differentiation were examined. Scanning electron microscopy (SEM), quantitative real times PCR (qPCR), Immunostaining and flow cytometry were used to analyze fabricated scaffold and the presence of IPCs. Functional maturity of differentiated cells was determined by measuring insulin release and the creation of IPCs was confirmed via gene and protein expression. In this study, the induced CJMSCs were morphologically similar to pancreatic islet-like cells. The expression of the islet-associated genes (glucagon, insulin and Pdx-1) and the insulin release (2.5-fold) in 3D-cultured cells was significantly higher than the 2D. The expression of IPCs genes was significantly higher in CJMSCs differentiated on plasma-treated nanofibers compared to those on untreated scaffolds. In conclusion, the results show that CJMSCs might be a new source for Diabetic Mellitus therapy and the nanofibrous scaffold could be used as a potential cell carrier for islet tissue engineering.

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Isolation and Expansion of Mesenchymal Stem Cells from Human Conjunctival Tissue

Cited by 3 publications

References 24 publications

Fibrin gel as a scaffold for photoreceptor cells differentiation from conjunctiva mesenchymal stem cells in retina tissue engineering

Fibrin gel as a scaffold for photoreceptor cells differentiation from conjunctiva mesenchymal stem cells in retina tissue engineering

Effect of parameters on the quality of core‐shell fibrous scaffold for retinal differentiation of conjunctiva mesenchymal stem cells

Differentiation of conjunctiva mesenchymal stem cells into secreting islet beta cells on plasma treated electrospun nanofibrous scaffold

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