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1.
Patcharee Prasajak Piyaporn Rattananinsruang Kamonnaree Chotinantakul Chavaboon Dechsukhum Wilairat Leeanansaksiri 《Cytotechnology》2015,67(3):493-505
Mesenchymal stem cells (MSCs) are accepted as a promising tool for therapeutic purposes. However, low proliferation and early senescence are still main obstacles of MSCs expansion for using as cell-based therapy. Thus, clinical scale of cell expansion is needed to obtain a large number of cells serving for further applications. In this study, we investigated the value of embryonic stem cells conditioned medium (ESCM) for in vitro expansion of Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) as compared to typical culture medium for MSCs, Dulbecco’s modified Eagle’s medium with 1.0 g/l glucose (DMEM-LG) supplemented with 10 % FBS, under hypoxic condition. The expanded cells from ESCM (ESCM-MSCs) and DMEM-LG (DMEM-MSCs) were characterized for both phenotype and biological activities including proliferation rate, population doubling time, cell cycle distribution and MSCs characteristics. ESCM and DMEM-LG could enhance WJ-MSCs proliferation as 204.66 ± 10.39 and 113.77 ± 7.89 fold increase at day 12, respectively. ESCM-MSCs could express pluripotency genes including Oct-4, Oct-3/4, Nanog, Klf-4, C-Myc and Sox-2 both in early and late passages whereas the downregulations of Oct-4 and Nanog were detected in late passage cells of DMEM-MSCs. The 2 cell populations also showed common MSCs characteristics including normal cell cycle, fibroblastic morphology, cell surface markers expressions (CD29+, CD44+, CD90+, CD34−, CD45−) and differentiation capacities into adipogenic, chondrogenic and osteogenic lineages. Moreover, our results revealed that ESCM exhibited as a rich source of several factors which are required for supportive WJ-MSCs proliferation. In conclusion, ESCM under hypoxic condition could accelerate WJ-MSCs expansion while maintaining their pluripotency properties. Our knowledge provide short term and cost-saving in WJ-MSCs expansion which has benefit to overcome insufficient cell numbers for clinical applications by reusing the discarded cell culture supernates from human ES culture system. Moreover, these findings can also apply for stem cell banking, regenerative medicine and pharmacological applications. 相似文献
2.
Recently, stem cell biology has become an interesting topic, especially in the context of treating diseases and injuries using
transplantation therapy. Several varieties of human stem cells have been isolated and identified in vivo and in vitro. Ideally, stem cells for regenerative medical application should be found in abundant quantities, harvestable in a minimally
invasive procedure, then safely and effectively transplanted to either an autologous or allogenic host. The two main groups
of stem cells, embryonic stem cells and adult stem cells, have been expanded to include perinatal stem cells. Mesenchymal
stem cells from perinatal tissue may be particularly useful in the clinic for autologous transplantation for fetuses and newborns,
and after banking in later stages of life, as well as for in utero transplantation in case of genetic disorders. 相似文献
3.
In vitro chondrogenesis of Wharton’s jelly mesenchymal stem cells in hyaluronic acid-based hydrogels
Ewelina Aleksander-Konert Piotr Paduszyński Alicja Zajdel Zofia Dzierżewicz Adam Wilczok 《Cellular & molecular biology letters》2016,21(1):11
Background
In this study, we evaluated the usefulness of two commercially available hyaluronic acid-based hydrogels, HyStem and HyStem-C, for the cultivation of Wharton’s jelly mesenchymal stem cells (WJ-MSCs) and their differentiation towards chondrocytes.Methods
The WJ-MSCs were isolated from umbilical cord Wharton’s jelly using the explant method and their immunophenotype was evaluated via flow cytometry analysis. According to the criteria established by the International Society for Cellular Therapy, they were true MSCs. We assessed the ability of the WJ-MSCs and chondrocytes to grow in three-dimensional hydrogels and their metabolic activity. Chondrogenesis of WJ-MSCs in the hydrogels was determined using alcian blue and safranin O staining and real-time PCR evaluation of gene expression in the extracellular matrixes: collagen type I, II, III and aggrecan.Results
Chondrocytes and WJ-MSCs cultured in the HyStem and HyStem-C hydrogels adopted spherical shapes, which are characteristic for encapsulated cells. The average viability of the WJ-MSCs and chondrocytes in the HyStem hydrogels was approximately 67 % when compared with the viability in 2D culture. Alcian blue and safranin O staining revealed intensive production of proteoglycans by the cells in the HyStem hydrogels. Increased expression of collagen type II and aggrecan in the WJ-MSCs cultured in the HyStem hydrogel in the presence of chondrogenic medium showed that under these conditions, the cells have a high capacity to differentiate towards chondrocytes. The relatively high viability of WJ-MSCs and chondrocytes in both HyStem hydrogels suggests the possibility of their use for chondrogenesis.Conlusions
The results indicate that WJ-MSCs have some degree of chondrogenic potential in HyStem and HyStem-C hydrogels, showing promise for the engineering of damaged articular cartilage.4.
Immunological impact of Wharton’s Jelly mesenchymal stromal cells and natural killer cell co-culture
Mehdi Najar Mohammad Fayyad-Kazan Nathalie Meuleman Dominique Bron Hussein Fayyad-Kazan Laurence Lagneaux 《Molecular and cellular biochemistry》2018,438(1-2):111-121
In palmipeds, overfeeding leads to hepatic steatosis, also called “foie gras” which is the result of many metabolic mechanisms. In order to understand these mechanisms, we decided to measure the expression of genes implicated in lipid metabolism during 12 hours (h) following the last meal of the overfeeding period. We have shown that there is a precocious expression (within 2 h) of fatty acid synthase and acyl CoA synthetase long-chain 1 in liver and muscle of mule ducks in addition with a later peak. Furthermore, di-acyl glycerol acyl transferase presents the highest induction of expression in liver and it is overexpressed quite a long time, positioning this enzyme as a key factor in hepatic steatosis. These observations are quite similar in muscle. Lipoprotein secretion is upregulated later in postprandial period, with an upregulation of apolipoprotein and microsomal triglycerides transfer protein beginning at 5 h in liver or muscle. Regarding hepatic re-uptake of lipid, lesser variations are observed, suggesting that fatty acid binding protein and very low-density lipoprotein receptor (VLDLR) are already at their maximum expression specifically in these tissues. In muscle, VLDLR and LDLR upregulation is observed 5 h after the meal, associated with an overexpression in the adipose tissue of lipase maturation factor 1 involved in the maturation of lipoprotein lipase. These findings will allow us to better understand the kinetic treatment in lipid metabolism after a meal in overfed ducks. This first report on kinetic expression will allow researcher to better target their sampling time knowing the optimal point of expression of each gene. 相似文献
5.
Salehinejad P Alitheen NB Ali AM Omar AR Mohit M Janzamin E Samani FS Torshizi Z Nematollahi-Mahani SN 《In vitro cellular & developmental biology. Animal》2012,48(2):75-83
Several techniques have been devised for the dissociation of tissues for primary culture. These techniques can affect the
quantity and quality of the isolated cells. The aim of our study was to develop the most appropriate method for the isolation
of human umbilical cord-derived mesenchymal (hUCM) cells. In the present study, we compared four methods for the isolation
of hUCM cells: three enzymatic methods; collagenase/hyaluronidase/trypsin (CHT), collagenase/trypsin (CT) and trypsin (Trp),
and an explant culture (Exp) method. The trypan blue dye exclusion test, the water-soluble tetrazolium salt-1 (WST-1) assay,
flow cytometry, alkaline phosphatase activity and histochemical staining were used to evaluate the results of the different
methods. The hUCM cells were successfully isolated by all methods but the isolation method used profoundly altered the cell
number and proliferation capacity of the isolated cells. The cells were successfully differentiated into adipogenic and osteogenic
lineages and alkaline phosphatase activity was detected in the hUCM cell colonies of all groups. Flow cytometry analysis revealed
that CD44, CD73, CD90 and CD105 were expressed in all groups, while CD34 and CD45 were not expressed. The expression of C-kit
in the enzymatic groups was higher than in the explant group, while the expression of Oct-4 was higher in the CT group compared
to the other groups. We concluded that the collagenase/trypsin method of cell isolation yields a higher cell density than
the others. These cells expressed a higher rate of pluripotent cell markers such as C-kit and Oct-4, while the explant method
of cell isolation resulted in a higher cell proliferation rate and activity compared to the other methods. 相似文献
6.
Fatemeh Hendijani Hojjat Sadeghi-Aliabadi Shaghayegh Haghjooy Javanmard 《Cell and tissue banking》2014,15(4):555-565
Adult stem cells are of particular importance for applications in regenerative medicine. Umbilical cord was established recently as an alternative source of mesenchymal stem cell (MSC) instead of bone marrow (BM) and is superior to BM and other adult tissues according to several MSC properties. Additionally, for the purpose of cell therapy in clinical scale, steps of cell isolation, expansion and culture required to be precisely adjusted in order to obtain the most cost-effective, least time-consuming, and least labor-intensive method. Therefore, in this study, we are going to compare two simple and cost-effective explant culture methods for isolation of MSCs from human umbilical cord. One of the methods isolates cells from entire cord and the other from Wharton’s jelly matrix. Isolated cells then cultured in simple medium without addition of any growth factor. MSCs obtained via both methods display proper and similar characteristics according to morphology, population doubling time, post-thaw survival, surface antigenicity and differentiation into adipocytes, osteocytes, and chondrocytes. MSCs can easily be obtained from the entire cord and Wharton’s jelly, and it seems that both tissues are appropriate sources of stem cells for potential use in regenerative medicine. However, from technical largescale preview, MSC isolation from entire cord piece is less labor-intensive and time-consuming than from Wharton’s jelly part of the cord. 相似文献
7.
8.
《European journal of cell biology》2022,101(3):151245
Mesenchymal stem cells (MSCs) show a decline in pluripotency and differentiation with increased cell culture passages in 2D cultures. The 2D monolayer culture fails to correctly imitate the architecture and microenvironments of in-vivo cell models. Alternatively, 3D culture may improve the simulations of in-vivo cell microenvironments with wide applications in cell culture and drug discovery. In the present study, we compared various 3D culturing techniques such as 3D micro-well (3D-S), hanging drop (HD), and ultra-low attachment (ULA) plate-based spheroid culture to study their effect on morphology, viability, pluripotency, cell surface markers, immunomodulatory factors, and differentiation capabilities of Wharton’s jelly-mesenchymal stem cells (WJ-MSCs). The cell morphology, viability, and senescence of 3D cultured WJ-MSCs were comparable to cells in 2D culture. The expression of pluripotency markers (OCT4, SOX2, and NANOG) was enhanced upto 2–8 fold in 3D cultured WJ-MSCs when compared to 2D culture. Moreover, the immunomodulatory factors (IDO, IL-10, LIF, ANG1, and VEGF) were significantly elevated in ULA based 3D cultured WJ-MSCs. Furthermore, significant enhancement in the differentiation potential of WJ-MSCs towards adipocyte (ADP and C/EBP-α), osteocyte (OPN and RUNX2), and definitive endodermal (SOX17, FOXA2, and CXCR4) lineages in 3D culture conditions were observed. Additionally, the osteogenic and adipogenic differentiation potential of WJ-MSCs over the time points 7 days, 14 days, and 28 days was also significantly increased in 3D culture groups. Our study demonstrates that stemness properties of WJ-MSCs were significantly enhanced in 3D cultures and ULA-based culture outperformed other methods with high pluripotency gene expression and enhanced differentiation potential. This study indicates the efficacy of 3D cultures to bridge the gap between 2D cell culture and animal models in regenerative medicine. 相似文献
9.
Karaca Cigdem Bostancıeri Nuray Ovayolu Ali Kahraman Demet Tasdemir 《Molecular biology reports》2020,47(12):9313-9323
Molecular Biology Reports - The current study investigated the change in umbilical cord tissue and the number of markers of Wharton’s jelly mesenchymal stem cells (WJ-MSC) in pregnant women... 相似文献
10.
Hu CC Ke YQ Sun XL Jiang XD Xu RX Lv J Wang YS Cai YQ Qin LS Zou YX 《Biotechnology letters》2009,31(2):181-189
Human mesenchymal stem cells-like cells (hMSCs-like cells) were used as a tumor treatment platform for the systemic delivery
of immunotoxin genes. VEGF165-PE38 recombinant immunotoxin served as the model system. hMSCs-like cells were isolated, expanded,
and electroporated with the pIRES2-VEGF165PE38-EGFP plasmid. RT-PCR and ELISA were used to confirm the expression of VEGF165-PE38
in the transfected hMSCs-like cells. These cells released 1390 ± 137 pg VEGF165-PE38/104cells over 48 h into the culture medium and the supernatant was capable of selectively killing human umbilical vein endothelial
cells (HUVECs) and increasing apoptosis in these cells. In contrast, RPMI8226 was not inhibited by identical supernatants.
Thus, these results lay the foundation for further studies on the potential role of hMSCs-like cells as a targeted therapeutic
delivery vehicle for immunotoxins.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
11.
Kun Liu Wei Zhang Yunlong Li Qingfeng Ding Yunan Bai Fachen Wang Guangming Xu 《Cytotechnology》2018,70(5):1291-1299
The potency of mesenchymal stem cells (MSCs) for tissue repair and regeneration is mainly based on their ability to secret beneficial molecules. Administration of MSCs has been proposed as an innovative approach and is proved by a number of clinical trials to a certain degree for the therapy of many diseases including Parkinson’s disease (PD). However, the efficacy of MSCs alone is not significant. We investigated the effect of neurotrophic tyrosine receptor kinase 1 (NTRK1) overexpressed peripheral blood MSCs (PB-MSCs) on PD rat model. NTRK1 was overexpressed in PB-MSCs, which were then injected into PD rat model, Dopaminergic (DA) neuron regeneration and rotational performance was assessed. We found that DA neuron repair was increased in lesion site, rotational performance was also improved in MSC transplanted PD rat, with most potent effect in NTRK1 overexpressed PB-MSC transplanted PD rat. Our results indicate that overexpression of NTRK1 in MSCs could be an optimized therapeutic way via MSCs for PD treatment. 相似文献
12.
Isolation and characterization of mesenchymal stem cells derived from bone marrow of patients with Parkinson’s disease 总被引:1,自引:0,他引:1
Mesenchymal stem cells (MSCs) are capable of self-renewing and differentiating into multiple tissues; they are expected to become a source of cells for regenerative therapy. Compared to allogeneic MSCs, autologous MSCs from patients needing cell-based therapy may be an ideal alternative stem cell source. However, characterizations of MSCs from a disease state remains extremely limited. Therefore, we have isolated and characterized MSCs from Parkinson's disease (PD) patients and compared them with MSCs derived from normal adult bone marrow. Our results show that PD-derived MSCs are similar to normal MSCs in phenotype, morphology, and multidifferentiation capacity. Moreover, PD-derived MSCs are capable of differentiating into neurons in a specific medium with up to 30% having the characteristics of dopamine cells. At last, PD-derived MSCs could inhibit T-lymphocyte proliferation induced by mitogens. These findings indicate that MSCs derived from PD patients' bone marrow may be a promising cell type for cellular therapy and somatic gene therapy applications. 相似文献
13.
GUAN FangXia MA ShanShan SHI XinYi MA Xun CHI LianKai LIANG Shuo CUI YuanBo WANG ZhiBin YAO Ning GUAN ShaoKang YANG Bo 《中国科学:生命科学英文版》2014,57(2):181-187
Seeding cells and scaffolds play pivotal roles in bone tissue engineering and regenerative medicine.Wharton’s jelly-derived mesenchymal stem cells(WJCs)from human umbilical cord represent attractive and promising seeding cells in tissue regeneration and engineering for treatment applications.This study was carried out to explore the biocompatibility of scaffolds to seeding cells in vitro.Rod-like nano-hydroxyapatite(RN-HA)and flake-like micro-hydroxyapatite(FM-HA)coatings were prepared on Mg-Zn-Ca alloy substrates using micro-arc oxidation and electrochemical deposition.WJCs were utilized to investigate the cellular biocompatibility of Mg-Zn-Ca alloys after different surface modifications by observing the cell adhesion,morphology,proliferation,and osteoblastic differentiation.The in vitro results indicated that the RN-HA coating group was more suitable for cell proliferation and cell osteoblastic differentiation than the FM-HA group,demonstrating better biocompatibility.Our results suggested that the RN-HA coating on Mg-Zn-Ca alloy substrates might be of great potential in bone tissue engineering. 相似文献
14.
15.
Differentiation profile of brain tumor stem cells: a comparative study with neural stem cells 总被引:25,自引:0,他引:25
Understanding of the differentiation profile of brain tumor stem cells (BTSCs), the key ones among tumor cell population, through comparison with neural stem cells (NSCs) would lend insight into the origin of glioma and ultimately yield new approaches to fight this intractable disease. Here, we cultured and purified BTSCs from surgical glioma specimens and NSCs from human fetal brain tissue, and further analyzed their cellular biological behaviors, especially their differentiation property. As expected, NSCs differentiated into mature neural phenotypes. In the same differentiation condition, however, BTSCs exhibited distinguished differences. Morphologically, cells grew flattened and attached for the first week, but gradually aggregated and reformed floating tumor sphere thereafter. During the corresponding period, the expression rate of undifferentiated cell marker CD 133 and nestin in BTSCs kept decreasing, but 1 week later, they regained ascending tendency. Interestingly, the differentiated cell markers GFAP and β-tubulinlII showed an expression change inverse to that of undifferentiated cell markers. Taken together, BTSCs were revealed to possess a capacity to resist differentiation, which actually represents the malignant behaviors of glioma. 相似文献
16.
Parkinson’s disease (PD) is the most common movement disorder. The neuropathology is characterized by the loss of dopamine
neurons in the substantia nigra pars compacta. Transplants of fetal/embryonic midbrain tissue have exhibited some beneficial
clinical effects in open-label trials. Neural grafting has, however, not become a standard treatment for several reasons.
First, the supply of donor cells is limited, and therefore, surgery is accompanied by difficult logistics. Second, the extent
of beneficial effects has varied in a partly unpredictable manner. Third, some patients have exhibited graft-related side
effects in the form of involuntary movements. Fourth, in two major double-blind placebo-controlled trials, there was no effect
of the transplants on the primary endpoints. Nevertheless, neural transplantation continues to receive a great deal of interest,
and now, attention is shifting to the idea of using stem cells as starting donor material. In the context of stem cell therapy
for PD, stem cells can be divided into three categories: neural stem cells, embryonic stem cells, and other tissue-specific
types of stem cells, e.g., bone marrow stem cells. Each type of stem cell is associated with advantages and disadvantages.
In this article, we review recent advances of stem cell research of direct relevance to clinical application in PD and highlight
the pros and cons of the different sources of cells. We draw special attention to some key problems that face the translation
of stem cell technology into the clinical arena. 相似文献
17.
Vaishnevi Raj Stephanie Claudine Arjunan Subramanian Kimberley Tam Arijit Biswas Ariff Bongso Chui-Yee Fong 《Journal of cellular biochemistry》2019,120(7):11222-11240
We showed in previous studies that human umbilical cord Wharton’s jelly stem cells (hWJSCs) improved the healing rates of excisional and diabetic wounds in the mouse model. As an extension of those studies, we report here the more detailed quantitative histological, immunohistochemical, and genomic evaluation of biopsies from those excisional and diabetic wounds in an attempt to understand the mechanisms of the enhanced wound healing aided by hWJSCs. Bright-field microscopic observations and ImageJ software analysis on histological sections of the excisional and diabetic wound biopsies collected at different time points showed that the thickness of the epidermis and dermis, and positive picrosirius-red stained areas for collagen, were significantly greater in the presence of hWJSCs compared with controls (P < 0.05). Immunohistochemistry of the diabetic wound biopsies showed increased positive staining for the vascular endothelial marker CD31 and cell proliferation marker Ki67 in the presence of hWJSCs and its conditioned medium (hWJSC-CM). Quantitative real-time polymerase chain reaction showed upregulation of groups of genes involved in extracellular matrix regulation, collagen biosynthesis, angiogenesis, antifibrosis, granulation, and immunomodulation in the presence of hWJSCs. Taken together, the results demonstrated that hWJSCs and hWJSC-CM that contains the paracrine secretions of hWJSCs, enhance the healing of excisional and diabetic wounds via re-epithelialization, collagen deposition, angiogenesis, and immunomodulation. The inclusion of an Aloe vera-polycaprolactone (AV/PCL) nanocarrier did not significantly change the effect of the hWJSCs. However, the topical application of an AV/PCL nanocarrier impregnated with hWJSCs is convenient and less invasive than the administration of hWJSC injections into wounds. 相似文献
18.
19.
Lili Du Runxiao Lv Xiaoyi Yang Shaohang Cheng Jing Xu Tingxian Ma 《Biotechnology letters》2016,38(6):931-939
Objectives
To explore the effect of placenta-derived mesenchymal stem cells on scar formation as well as the underlying mechanism.Results
The isolated placenta-derived mesenchymal stem cells from mice were distributed in the wounded areas of scalded mouse models, attenuated inflammatory responses and decreased the deposition of collagens, thus performing a beneficial effect against scar formation. Hypoxia enhanced the protective effect of placenta-derived mesenchymal stem cells and hypoxia-inducible factor-1α was involved in the protective effect of placenta-derived mesenchymal stem cells in hypoxic condition.Conclusions
Hypoxia enhanced the protective effect of placenta-derived mesenchymal stem cells through hypoxia-inducible factor-1α and PMSCs may have a potential application in the treatment of wound.20.
C-K Yang C-C Feng J-F Lo J-W Chen V. V Padma C-H Lai 《Biotechnic & histochemistry》2013,88(8):632-639
Mesenchymal stem cells are an attractive source of multipotent cells in part because they are easy to obtain. Several E3 ligases regulate the stability and functions of various factors in different adult stem cells through the ubiquitylation pathway. We investigated the C-terminus of Hsc70-interacting protein (CHIP) E3 ligase that regulates pluripotency of human Wharton’s jelly mesenchymal stem cells (hWJMSC). We found that CHIP increases protein kinase B (Akt) phosphorylation by decreased expression of phosphatase and tensin homolog (PTEN), which suggests improvement of the survival pathway by CHIP over-expression. We also found that increased CHIP expression induced Sox2 and NANOG, which can promote stem cell self-renewal and prevent oxidative stress-induced senescence of hWJMSC by decreased p21. We found that CHIP could be used to enhance the multiple functions of hWJMSC. 相似文献