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Bronchopulmonary dysplasia (BPD) is a chronic lung disease due to impaired pulmonary development and is one of the main causes of respiratory failure in preterm infants. Preterm infants with BPD have significantly higher complication and mortality rates than those without BPD. At present, comprehensive management is the main intervention method for BPD, including reasonable respiratory and circulatory support, appropriate enteral nutrition and parenteral nutrition, application of caffeine/glucocorticoids/surfactants, and out-of-hospital management after discharge. The continuous advances in stem cell medicine in recent years provide new ideas for the treatment of BPD. Various pre-clinical trials have confirmed that stem cell therapy can effectively prevent lung injury and promote lung growth and damage repair. This article performs a comprehensive analysis of the mechanism of mesenchymal stem cells in the treatment of BPD, so as to provide a basis for clinical applications.
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Humans , Infant, Newborn , Bronchopulmonary Dysplasia/prevention & control , Enteral Nutrition , Infant, Premature , Lung , Mesenchymal Stem CellsABSTRACT
BACKGROUND: Studies have shown that mesenchymal stem cells can reduce inflammation, promote wound healing, and reduce scar formation in wound healing. However, previous two-dimensional culture environment can lead to differences in gene expression, signal transduction, and morphology because of intracellular contact inhibition. OBJECTIVE: To investigate whether the ability of wound healing related factors secreted by human amniotic mesenchymal stem cells is affected by two-dimensional or three-dimensional culture environment. METHODS: The human amniotic mesenchymal stem cells cultured by traditional enzyme digestion method were inoculated in traditional cell culture flask (two-dimensional culture group) and ShakeGelTM 3D hydrogel (three-dimensional culture group) and induced to differentiate into adipocytes, osteoblasts, and chondrocytes, respectively. The direction of cell differentiation was determined by immunofluorescence staining. Human amniotic mesenchymal stem cells were fused to 70%-80% in two culture environments, and the growth characteristics and morphology of cells were observed under inverted phase contrast microscope and laser confocal microscope. After 24 hours of culture, relative mRNA expression of wound healing-related factors was detected by reverse transcription-quantitative polymerase chain reaction. After 48 hours of culture, the protein expression of wound healing-related factors was detected by the enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: (1) Human amniotic mesenchymal stem cells cultured in two-dimensional culture group were flat and spindle-shaped, which was a typical mesenchymal stem cell-like morphology. Human amniotic mesenchymal stem cells in the three-dimensional culture group were round and evenly dispersed in each layer of the hydrogel. (2) Human amniotic mesenchymal stem cells in the three-dimensional culture group exhibited the potential to differentiate into adipocytes, osteoblasts, and chondrocytes. (3) The mRNA expression of interleukin-6, interleukin-8, epidermal growth factor, basic fibroblast growth factor, hyaluronic acid, hepatocyte growth factor, and vascular endothelial growth factor in the three-dimensional culture group was significantly higher than that in the two-dimensional culture group (P 0.05). (4) The protein expression of interleukin-6, interleukin-8, interleukin-10, epidermal growth factor, basic fibroblast growth factor, hepatocyte growth factor, transforming growth factor β1 and vascular endothelial growth factor in the three-dimensional culture group was significantly higher than that in the two-dimensional culture (P 0.05). (5) These findings suggest that human amniotic mesenchymal stem cells cultured in three-dimensional hydrogel show better morphology and more encouraging paracrine effect of wound healing related factors than those cultured in traditional two-dimensional culture environment.
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BACKGROUND: Spinal cord injury is one of the diseases with highest disability. Due to the limited regeneration ability of spinal axons, it is difficult to recover and often leads to severe neurological sequelae. Treatment for spinal cord injury has become a hotspot. With the continuous development of biological and molecular biology research, mesenchymal stem cells have been found to possess big potential for treating spinal cord injury, and bring hope for the repair of spinal cord injury. OBJECTIVE: To review the treatment efficacy, property, application, limitations and prospects of mesenchymal stem cells in the treatment of spinal cord injury. METHODS: Databases of PubMed, Web of Science, WanFang and CNKI were searched for the articles concerning mesenchymal stem cells in the treatment of spinal cord injury. The keywords were “spinal cord injury, mesenchymal stem cell” in English and Chinese, respectively. Finally, 85 articles eligible for the inclusion and exclusion criteria were included for result analysis. RESULTS AND CONCLUSION: Mesenchymal stem cells repair spinal cord injury through a variety of mechanisms. For example, after in vivo transplantation, mesenchymal stem cells differentiate into neurons to replace damaged neurons. They secrete various nutrients through paracrine to regulate the microenvironment of spinal cord injury. Meanwhile, mesenchymal stem cells produce various extracellular matrixes, and provide support for axon regeneration, thus promoting neuronal axon regeneration. Although basic experiments have shown that mesenchymal stem cells exert satisfactory role in the treatment of spinal cord injury, the clinical outcome of mesenchymal stem cells is unsatisfactory. It is necessary to further explore the mechanism of action of mesenchymal stem cells in spinal cord injury.
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Objective: To establish the approach of isolating, culturing and identifing endometrial stem cells (EnSCs) derived from ectopic lesion of endometriosis patient; and preliminarily examine the biological characteristic of ectopic EnSCs, which provide support for further study on the potential role of ectopic EnSCs in the pathogenesis of endometriosis. Methods: The ectopic lesions of endometriosis were harvested from the patients with the informed consent and transferred to lab as soon as possible. The ectopic lesions were minced, digested by collagenase I and seeded into cell culture flasks for conventional culture (n=10). Expression of vimentin in ectopic EnSCs was examined by immunofluorescence (n = 3). Proliferative capacity of ectopic EnSCs was examined by MTT assay (n = 5). Multilineage differentiation potential of ectopic EnSCs was examined by adipogenic and osteogenic differentiation respectively (n = 3). Immunophenotype analysis of ectopic EnSCs was determined by flow cytometry (n = 3). Production of biological factors in ectopic EnSCs derived conditional medium (n = 6) and expression of adhesion molecules on ectopic EnSCs (n = 7) were examined by protein assays. Results: We successfully isolated EnSCs from ectopic lesions of endometriosis patients, and ectopic EnSCs were positive for vimentin and typical markers of mesenchymal stem cell (CD29, CD73, CD90 and CD105), and negative for the markers of hematopoietic stem cell (CD34 and CD45). The induced ectopic EnSCs showed obvious lipid droplets (adipogenic differentiation) and calcium nodules (osteogenic differentiation). The ectopic EnSCs could secrete high concentration of angiogenic factors [vascular endothelial growth factor (VEGF), angiotensin (ANG) and platelet-derived growth factor (PDGF)-AA]and angiogenesis associated inflammation cytokines [interleukin (IL-6), IL-8 and monocyte chemotactic protein l(MCP-l)]. Additionally, adhesion molecules analysis demonstrated the high expression of activated leukocyte adhesion molecule (ACLAM) and intercellular cell adhesion molecule-1 (ICAM-1) on ectopic EnSCs. Conclusion: We successfully establish the procedure of isolating and culturing ectopic EnSCs and demonstrate that ectopic EnSCs is capable of promoting angiogenesis through secreting high concentration of associated biological factors. The above result confirm the existence of EnSCs in ectopic lesions of endometriosis, which not only supports the stem cell based pathogenesis of endometriosis, but also shows the therapeutic potential of taking ectopic EnSCs as promising targets in the treatment of endometriosis.
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Age-related macular degeneration (AMD) is the major cause of blindness in the elderly, which mainly affects the retinal pigment epithelial (RPE) cells and photoreceptors, and progressively leads to the loss of central vision.AMD can be divided into dry AMD and wet AMD.At present, the treatment of AMD mainly aims at choroidal neovascularization (CNV) associated with wet AMD.However, there is no effective treatment for dry AMD.The common treatment methods include photodynamic therapy (PDT) and intravitreal injection of anti-vascular endothelial growth factor (VEGF). Furthermore, there are also laser therapy, radiotherapy and surgical treatment.However, at this stage, the clinical treatment methods can not eradicate the causes, and some patients show poor response to treatments.So people began to explore new treatments.Mesenchymal stem cells (MSCs) therapy has the characteristics of self-renewal and multidirectional differentiation potential, lower immunogenicity, easier access to materials and less ethical constraints, showing a broad therapeutic prospect.At present, many studies on the treatment of AMD with MSCs have been carried out at home and abroad, with fruitful results.This article elaborated the classification and treatment status of AMD and their limitations and summarized the research results of MSCs in the treatment of AMD at home and abroad in recent years.
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Objective: To review the advances in utilizing paracrine effect of stem cells in knee osteoarthritis (OA) treatment. Methods: The researches in applying stem cells derived conditioned medium, extracellular matrix, exosomes, and microvesicles in knee OA treatment and cartilage repair were reviewed and analyzed. Results: The satisfying outcomes of using different products of stem cells paracrine effect in knee OA condition as well as cartilage defect is revealed in studies in vitro and in vivo. The mechanism including suppressing the intraarticular inflammation, the apoptosis of chondrocytes, and the degradation of cartilage matrix, while enhancing the synthesis of cartilage matrix, the differentiation of in-situ stem cells into chondrocytes and the migration to the affected area. The effectiveness can be further improved supplemented with the tissue engineering methods or gene modification. Conclusion: Compared with the traditional stem cell therapy, applying the products from paracrine effect of stem cells in knee OA treatment is more economical and safer, presenting great potential in clinical practice.
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AIM:To investigate the regulation ofβ-adrenergic receptor (β-AR) agonist isoproterenol (ISO) on cardiac microRNA-21 (miR-21) expression.METHODS:The primary cultured mouse cardiomyocytes and cardiac fibroblasts were isolated by enzyme digestion and treated with ISO at 10μmol/L for 1, 6, 12, 24 and 48 h.The expression of miR-21 was detected by real-time PCR.The protein levels of p-STAT3 and STAT3 were determined by Western blot, and the concentration of interleukin-6 (IL-6) in the cultured supernatant was measured by ELISA.The cells were transfected with the luciferase reporter gene plasmid p GL3-21PPR containing the miR-21 promoter region, and the luciferase reporter gene assay was used to examine the effect of conditioned medium on the transcriptional activity of miR-21.RESULTS:The medium supernatant produced by ISO on cardiac fibroblasts was used as the conditioned medium, which increased the miR-21 expression in the cardiomyocytes in a time-dependent manner after fibroblasts was treated with ISO (P<0.05).The conditioned medium caused a significant increase in the transcriptional activity of miR-21 in the cardiomyocytes, while24 h and 48 h conditioned medium increased the transcriptional activity by 94.9％and 77.1％, respectively (P<0.01).The concentration of IL-6 in the conditioned medium was significantly increased, and the activity of transcriptional factor STAT3 was enhanced by paracrine action of IL-6 in the cardiomyocytes, which promoted the transcription and expression of miR-21.CONCLUSION:β-AR stimulation induces fibroblast synthesis and expression of IL-6 with paracrine effect on cardiomyocytes, up-regulates the expression of miR-21 in cardiomyocytes by IL-6/STAT3 pathway, and participates in the cardiac remodeling.
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Objective: To summarize the recent advances in the research of adipose-derived stem cells (ADSCs) for the treatment of refractory wounds.
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Objective To investigate the role and possible mechanisms of paracrine in radiation tolerance of non-small cell lung cancer (NSCLC) by observing the effects of radiation-resistant NSCLC H460R cell secretion on the radiosensitivity of parental H460 cells. Methods H460 cells were inoculated and cultured, and then divided into control group, conditional culturing group, irradiation group and irradiation combined conditional culturing group. One-time irradiation with 137Cs γ-rays was conducted with a dose rate of 1.02 Gy/min. Effect of H460R conditioned media on the radiation sensitivity of H460 cells was observed by clonogenic assay. The percentage of phospho-histone histone H2AX (γH2AX) positive cells was detected by immunofluorescence staining. The cyclical changes of the cells were detected by flow cytometry. The expression of DNA damage-associated proteinsγH2AX and Rad51 were detected by Western Blot. Results The results showed that compared with the singleγ-rays irradiation, the number of cell clones was significantly increased in the H460R cells after γ-rays irradiation combined with conditional culturing treatment at different doses of 2, 4, and 6 Gy, and the differences were statistically significant (2,4 Gy:all P<0.05;6 Gy:P<0.01). The results of immunofluorescence staining showed that the percentage ofγH2AX positive cells in the conditional culturing group was higher than that in the control group [(39.40±2.51)％vs. (25.21± 2.05)％], and the difference was statistically significant (P<0.01). This value in the irradiation combined conditional culturing group was lower than that in irradiated group [(60.48±2.79)％vs. (80.65±2.05)％], and the difference was statistically significant (P<0.001). The flow cytometry analysis showed that the percentage of cells in G2/M phase for the irradiation (4 Gy) combined conditional culturing group was higher than that of irradiated group [(26.83± 1.42)％ vs. (15.73±1.29)％], and the difference was statistically significant (P<0.001). The Western Blot results showed that γH2AX and Rad51 protein expression in the irradiation (4 Gy) combined conditional culturing group respectively was lower and higher than that in irradiated group. Conclusion In the tumor microenvironment, radiation-resistant H460R cells can enhance the radioresistance of H460 cells by paracrine, which may be related to the promotion of DNA repair ability after irradiation
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In addition to the impaired insulin secretion,the dysregulation of hyperglycermic hormone glucagon also takes part in the development and progress of diabetes,exacerbating the diabetic metabolic outcome.Although there has been a significant breakthrough in the current research on the regulation of insulin secretion,relatively little research on the roleof glucagon has been carried out.Actually,except for the paracrine regulation on glucagon secretion,pancreatic α-cell has also been proposed to be regulated by intrinsic mechanism,juxtacrine-mediated and sodium-glucose cotransporter.Furthermore,some factors outside the islet also affect on the glucagon secretion.Thus,more research on the complex secretory regulation of glucagon will provide a theoretical basis for our further understanding of the development of diabetes.In this review,we will summary the progress in the regulation of glucagon secretion in intra islets.
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Coculture between mesenchymal stem cells (MSCs) and chondrocytes has significant implications in cartilage regeneration. However, a conclusive understanding remains elusive. Previously, we reported that rabbit bone marrow-derived MSCs (rbBMSCs) could downregulate the differentiated phenotype of rabbit articular chondrocytes (rbACs) in a non-contact coculture system for the first time. In the present study, a systemic investigation was performed to understand the biological characteristics of chondrocytes in coculture with MSCs. Firstly, cells (MSCs and chondrocytes) from different origins were cocultured in transwell system. Different chondrocytes, when cocultured with different MSCs respectively, consistently demonstrated stimulated proliferation, transformed morphology and declined glycosaminoglycan secretion of chondrocytes. Next, cell surface molecules and the global gene expression of rbACs were characterized. It was found that cocultured rbACs showed a distinct surface molecule profile and global gene expression compared to both dedifferentiated rbACs and rbBMSCs. In the end, cocultured rbACs were passaged and induced to undergo the chondrogenic redifferentiation. Better growth and chondrogenesis ability were confirmed compared with control cells without coculture. Together, chondrocytes display comprehensive changes in coculture with MSCs and the cocultured rbACs are beneficial for cartilage repair.
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Cartilage , Chondrocytes , Chondrogenesis , Coculture Techniques , Gene Expression , Mesenchymal Stem Cells , Phenotype , Population Characteristics , RegenerationABSTRACT
This study aimed to explore the therapeutic effects of adipose-derived stem cells (ADSCs)-based microtissues (MTs) on erectile dysfunction (ED) in streptozotocin (STZ)-induced diabetic rats. Fifty-six 8-week-old Sprague-Dawley rats received intraperitoneal injection of STZ (60 mg kg-1 ), and 8 weeks later, the determined diabetic rats randomly received intracavernous (IC) injection of phosphate buffer solution (PBS), ADSCs, or MTs. Another eight normal rats equally got IC injection of PBS. MTs were generated with a hanging drop method, and the injected cells were tracked in ADSC- and MT-injected rats. Four weeks after the treatments, intracavernous pressure (ICP), histopathological changes in corpus cavernosum (CC), and functional proteins were measured. Rat cytokine antibody array was used to detect ADSCs or MTs lysate. The results showed that MTs expressed vascular endothelial growth factor (VEGF), nerve growth factor (NGF), and tumor necrosis factor-stimulated gene-6 (TSG-6). MTs injection had a higher retention than ADSCs injection and MTs treatment improved ICP, neuronal nitric oxide synthase (nNOS) expression, smooth muscle, and endothelial contents in diabetic rats, ameliorated local inflammation in CC better. Thus, our findings demonstrate that IC injection of MTs improves erectile function and histopathological changes in STZ-induced diabetic rats and appears to be more promising than traditional ADSCs. The underlying mechanisms involve increased cell retention accompanied with neuroprotection and anti-inflammatory behaviors of the paracrine factors.
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Liver fibrosis is a common chronic liver disease, which is a stress response process of liver cells affected by one or more pathogenies for long term or repeatedly. During the fibrosis process, massive accumulated extracellular matrix can form scar tissue, which results in liver dysfunction or failure and seriously endangers the health of people. According to many independent reports, stem cell therapy can facilitate the alleviation of liver fibrosis. During the stem cell therapy, stem cells migrate to the injury site of liver and alleviate the liver fibrosis by improving the microenvironment of the scar area via paracrine way. This article reviews the formation, treatment, and stem cell therapy of liver fibrosis.
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AIM: To investigate the therapeutic effect of endothelial progenitor cell-conditioned medium (EPC-CM) on the lung structure of neonatal rat exposed to hyperoxia, and to explore the mechanisms.METHODS: Bone marrow-derived endothelial progenitor cells (EPCs) were collected from new born Sprague-Dawley (SD) rats and the EPCs were identified.The conditioned medium from the passage 3 EPCs was collected.Newborn SD rats (n=40) were randomly divided into 4 groups.The rats in room air group were exposed to the room air (21% O2) for 21 d.The rats in hyperoxia group were exposed to hyperoxia (85% O2) for 21 d.The rats in endothelial cell basal medium (EBM) group were exposed to hyperoxia for 21 d, and received 100 μL EBM on postnatal day 14 (P14) in a single intratracheal (IT) injection.The rats in EPC-CM group were exposed to hyperoxia for 21 d, and received 100 μL EPC-CM on P14 in a singlie IT injection.The rats were sacrified on the 21st day.The left lungs were excised, placed in 4% paraformaldehyde, serially dehydrated in ethanol and embedded by paraffin.Serial sectioning of the paraffin-embedded left lung tissues was prepared for 5 μm thickness, and stained with hematoxylin and eosin.The pulmonary radical alveolar count (RAC) and alveolar mean linear intercept (MLI) were then calculated.The microvascular density was determined by FVIII immunostaining.The mRNA expression of KGF, VEGF, SP-A and SP-C in the right lung tissues was detected by real-time fluorescence quantitative PCR.RESULTS: The cultured cells had typical EPC morphological characteristics, and had the abilities to bind to FITC-UEA-1 and uptake DiI-ac-LDL.The body weight of the rats on day 21, RAC, MLI and microvascular density were significantly lower in hyperoxia group and EBM group than those in room air group (P<0.05).The EPC-CM group had significantly higher RAC and microvascular density than those in hyperoxia group and EBM group (P<0.05), but the body weight and MLI had no significant difference.The mRNA expression levels of KGF, VEGF, SP-A and SP-C in hyperoxia group and EBM group were significantly lower than those in room air group (P<0.05).The mRNA expression levels of KGF, VEGF, SP-A and SP-C in EPC-CM group were significantly higher than those in hyperoxia group and EBM group (P<0.05).CONCLUSION: EPC-CM promotes the lung alveolarization and microvascular formation in neonatal rats exposed to hyperoxia.These benefits may be correlated with the increased KGF and VEGF mRNA expression.
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Objective:Study on proliferation of gastric cancer cells (GCCs) and expressions of VEGF under the CO2 pneumoperitoneum environment,and effect of Bcl-2 specific inhibitor on the expression of VEGF in GCCs/TAMs co-culture system.Methods:TAMs induced by PMA and IL-4 in vitro.TAMs and MKN-45 cells were co-cultured in Transwell chamber,in pseudo CO2 pneumoperitoneum environment.Proliferation activity of GCCs was detected by MTT assay,and ELISA method was used to detect VEGF concentration in the cell culture supernatant.Results:Co-culture system was divided into CO2 pneumoperitoneum group and control group.Proliferation activity of MKN-45 cell and expression of VEGF of 5mmHg,10mmHg and 15mmHg groups are no difference with the control group;25 mmHg group is opposite to the former.The cells in co-culture group and CO2 pneumoperitoneum group (15 mmHg) were added to ABT-737,VEGF expression of co-culture + ABT-737 group was significantly lower than that of in the control group(P=0.001).Co-cultured cells VEGF expression in pneumoperitoneum group was significantly lower than the control group without inhibitor (P=0.000).Conclusion:Normal laparoscopic pneumoperitoneum is no stimulation of tumor cell proliferation defects,And higher pneumoperitoneum pressure might increase the hypoxia status of tumor microenvironment,promote tumor malignant progression.The interaction between tumor cells and TAMs may be achieved by paracrine Bcl-2-VEGF loop.
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Objective To explore the expression of miroRNA-21 (miRNA-21) in myocardial fibroblasts stimulated by indoxyl sulfate (IS) and its role on paracrine factors of myocardial fibroblasts.Methods Myocardial fibroblasts which derived from C57BL/6J mice were divided into control group and IS group,and their expressions of miRNA-21 were detected by real time PCR after 48 h.MiRNA-21 inhibitor transfection was applied to silence miRNA-21 expression.Myocardial fibroblasts were divided into creatinine (Scr) group (Scr treated for 48 h),Scr+IS group (Scr and 50 μmol/L IS treated for 48 h),Scr+miRNA-21 inhibitor group (miRNA-21 inhibitor treated for 24 h and then Scr treated for 48 h) and Scr+miRNA-21 inhibitor+ IS group (miRNA-21 inhibitor pretreated for 24 h and then Scr and IS treated for 48 h).Enzyme-linked immunosorbent assay was performed to evaluate the expressions of interleukin-1 (IL-1),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α).Western blotting was applied to detect the expressions of transforming growth factor-β (TGF-β),matrix metalloproteinases 2 (MMP2),matrix metalloproteinases 9 (MMP9) and tissue inhibitor of metalloproteinases 1 (TIMP1).Results The expression of miRNA-21 was obviously increased in IS group than that in control group (P ＜ 0.01).Compared with those in Scr group,the expressions of IL-1,IL-6,TNF-α,TGF-β,MMP2 and MMP9 significantly increased (all P ＜ 0.05),while the expression of TIMP1 decreased (P ＜ 0.05).When the expression of miRNA-21 was inhibited,the expressions of IL-1,IL-6,TNF-α,TGF-β and MMP2 in Scr+miRNA-21 inhibitor+IS group significantly decreased than those in Scr+IS group (all P ＜0.05),and the expression of TIMP1 decreased further (P ＜ 0.05).Conclusions IS can promote the expression of miRNA-21 in myocardial fibroblasts.MiRNA-21 plays an important role in regulating inflammatory factors and pro-fibrogenic cytokines in myocardial fibroblasts.
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Understanding the crosstalk mechanisms between perivascular cells (PVCs) and cancer cells might be beneficial in preventing cancer development and metastasis. In this study, we investigated the paracrine influence of PVCs derived from human umbilical cords on the proliferation of lung adenocarcinoma epithelial cells (A549) and erythroleukemia cells (TF-1α and K562) in vitro using Transwell® co-culture systems. PVCs promoted the proliferation of A549 cells without inducing morphological changes, but had no effect on the proliferation of TF-1α and K562 cells. To identify the factors secreted from PVCs, conditioned media harvested from PVC cultures were analyzed by antibody arrays. We identified a set of cytokines, including persephin (PSPN), a neurotrophic factor, and a key regulator of oral squamous cell carcinoma progression. Supplementation with PSPN significantly increased the proliferation of A549 cells. These results suggested that PVCs produced a differential effect on the proliferation of cancer cells in a cell-type dependent manner. Further, secretome analyses of PVCs and the elucidation of the molecular mechanisms could facilitate the discovery of therapeutic target(s) for lung cancer.
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Humans , Adenocarcinoma , Carcinoma, Squamous Cell , Coculture Techniques , Culture Media, Conditioned , Cytokines , Epithelial Cells , In Vitro Techniques , K562 Cells , Leukemia, Erythroblastic, Acute , Lung , Lung Neoplasms , Neoplasm Metastasis , Umbilical CordABSTRACT
Deregulation of Insulin like growth factors (IGF) is an important determinant of breast carcinogenesis. Circulatory IGF-1 (potent breast mitogen) and IGFBP-3 (its regulator) are extensively evaluated; few studies report transcript copy numbers (CN) from ex-vivo samples. This study from 106 patients evaluated mRNA expression (qRT-PCR CN) of IGF-1 and IGFBP-3 for prognostic and predictive utility from tumor, adjacent normal tissues (ANT) and lymph nodes. The differences in IGF-1 and IGFBP-3 mRNA levels (CN/μg RNA) were juxtaposed to clinical and pathologic variables and survival. Tumors expressed lower IGF-1 and higher IGFBP-3 as compared to ANT. Both transcript levels decreased with increasing age. Primary tumors with nodal involvement, Invasive Lobular carcinoma (ILC) histology and stromal involvement showed increased transcript levels than their respective counterparts. Moreover, surviving patients showing no relapse had higher expression of both molecules. Early stage and necrosed tumors expressed higher IGFBP-3 while a trend of lower expression was seen as tumor grade advanced. IGF-1 expression was inversely correlated to stage, histologic grade and. Significantly different Relapse Free Survival (RFS) was seen with IGFBP-3 up-/down-regulation considering progesterone receptor (PR) status but not estrogen receptor (ER) and HER-2 while the Overall Survival (OS) was similar for both these molecules. We conclude that expression of these molecules may aid prognostication and success of anti IGF-1 strategies.
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BACKGROUND: The stem cell-derived secretome has received considerable attention as an alternative to stem cells for therapeutic applications. However, establishing optimal culture conditions is key to obtaining appropriate secretome contents. Here, the optimal culturing environment for achieving a high-efficiency secretome was determined via hypoxic preconditioning of human adipose-derived stem cells (ASC). METHODS: Normoxic conditioned media (NCM) and hypoxic conditioned media (HCM) were obtained after culturing human ASCs under normoxia (20% O2) or hypoxia (1% O2), respectively. Subsequently, both normal and thioacetamide-induced hepatotoxic hepatocytes were treated with NCM or HCM. In addition, partially hepatectomized mice were infused with control saline, NCM, and HCM. The effects on liver regeneration and serum transaminases levels were then compared. RESULTS: Hypoxic preconditioning significantly increased mRNA expression of proinflammatory cytokines (interleukin-6 and tumor necrosis factor-α) and growth factors (hepatocyte growth factor and vascular endothelial growth factor). In both normal and thioacetamide-induced hepatotoxic hepatocyte (alpha mouse liver 12 [AML12]) cell lines, HCM treatment resulted in the highest cell viability (122% and 95%, respectively), followed by NCM (111% and 78%, respectively). In addition, intravenous administration of HCM to partially hepatectomized mice resulted in substantially enhanced liver regeneration compared with the NCM group (P<0.05). CONCLUSIONS: Taken together, the secretome obtained from ASC with hypoxic preconditioning showed potential to alleviate liver damage both in vitro and in vivo. Hypoxic culture of ASC is expected to play an important role in regenerative medicine by inducing secretome production that is beneficial for improving liver regeneration.
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Animals , Humans , Mice , Administration, Intravenous , Hypoxia , Cell Line , Cell Survival , Culture Media, Conditioned , Cytokines , Hepatocytes , Intercellular Signaling Peptides and Proteins , Liver Regeneration , Liver , Necrosis , Regenerative Medicine , RNA, Messenger , Stem Cells , TransaminasesABSTRACT
Diabetic nephropathy (DN) is one of the severe chronic complications of diabetes,and has become an important contributing factor of end-stage renal disease and increased mortality of diabetes.DN pathogenesis is complicated,involving many factors working together.Regenerative medicine based on stem cells for the treatment of DN brings new hope.The mechanism of mesenchymal stem cells (MSCs) improving DN development get more and more attention,in this paper,we will talk about MSCs in improving DN through paracrine effect,hope it will provide the new mentality for the treatment of DN.