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ABSTRACT: In this study, an injectable, biodegradable hydrogel composite of oligo[poly(ethylene glycol) fumarate] (OPF) was investigated as a carrier of mouse embryonic stem cells (mESCs) for the treatment of myocardial infarction (MI). The OPF hydrogels were used to encapsulate mESCs. The cell differentiation in vitro over 14 days was determined via immunohistochemical examination. Then, mESCs encapsulated in OPF hydrogels were injected into the LV wall of a rat MI model. Detailed histological analysis and echocardiography were used to determine the structural and functional consequences after 4 weeks of transplantation. With ascorbic acid induction, mESCs could differentiate into cardiomyocytes and other cell types in all three lineages in the OPF hydrogel. After transplantation, both the 24-hr cell retention and 4-week graft size were significantly greater in the OPF + ESC group than that of the PBS + ESC group (P < 0.01). Four weeks after transplantation, OPF hydrogel alone significantly reduced the infarct size and collagen deposition and improved the cardiac function. The heart function and revascularization improved significantly, while the infarct size and fibrotic area decreased significantly in the OPF + ESC group compared with that of the PBS + ESC, OPF and PBS groups (P < 0.01). All treatments had significantly reduced MMP2 and MMP9 protein levels compared to the PBS control group, and the OPF + ESC group decreased most by Western blotting. Transplanted mESCs expressed cardiovascular markers. This study suggests the potential of a method for heart regeneration involving OPF hydrogels for stem cell encapsulation and transplantation.
Journal of Cellular and Molecular Medicine 05/2012; 16(6):1310 - 1320. · 4.13 Impact Factor
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Zhiqiang Liu,
Haibin Wang,
Yan Wang,
Qiuxia Lin,
Anning Yao,
Feng Cao, Dexue Li,
Jin Zhou,
Cuimi Duan,
Zhiyan Du,
Yanmeng Wang,
Changyong Wang
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ABSTRACT: One challenge of cellular cardiomyoplasty for myocardial infarction (MI) is how to improve MI microenvironment to facilitate stem cell engraftment, survival and homing for myocardial repair. The application of injectable hydrogels is an effective strategy. However, it has not been thoroughly investigated on the role of the injectable scaffolds, in improving MI microenvironment, providing space and guidance for cell survival, engraftment and homing. We explored an injectable chitosan hydrogel for stem cell delivery into ischemic heart and investigated the beneficial effects and mechanisms of the hydrogel. In vitro, H(2)O(2)-treatment was used to mimic reactive oxygen species (ROS) microenvironment. The influence of ROS and protection of chitosan components on adipose-derived mesenchymal stem cells (ADSCs) was analyzed too. In vivo, MI was induced by the left anterior descending artery ligation in SD rats. PBS, chitosan hydrogel, ADSC/PBS and ADSC/chitosan hydrogel were injected into the border of infracted hearts respectively. Multi-techniques were used to assess the beneficial effects of chitosan hydrogel after transplantation. We observed that ROS generated by ischemia would impair ADSC adhesion molecules, including integrin-related adhesion molecules integrin αV and β1, focal adhesion-related molecules p-FAK and p-Src, and corresponding ligands of host myocardium ICAM1 and VCAM1. Chitosan hydrogel could rescue these molecules through ROS scavenging and recruit key chemokine for stem cell homing, such as SDF-1. The results suggest that chitosan hydrogel could improve MI microenvironment, enhance stem cell engraftment, survival and homing in ischemic heart through ROS scavenging and chemokine recruitment, contributing to myocardial repair.
Biomaterials 04/2012; 33(11):3093-106. · 7.40 Impact Factor
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ABSTRACT: OP (octylphenol), an environmental oestrogen was administered, and differentially expressed proteins were analysed in mice testes to clarify its mechanism of action in male sterility. Male Kunming suckling mice (10 days old) were subcutaneously injected with OP at a dose of 10 μg/kg per day, 50 μg/kg per day and 100 μg/kg per day as low-, medium- and high-dose groups, respectively, for 35 days. Animals in the control group received subcutaneous injections of olive oil at a dose of 10 μl/mouse per day. Serum oestradiol, testosterone, FSH (follicle stimulating hormone) and LH (luteinizing hormone) levels were measured on day 45. The left testes were removed for tissue analysis, and the right testes were analysed for differentially expressed proteins by using two-dimensional gel electrophoresis and MS. Tissue analysis showed that mice spermatogenesis was blocked at the round spermatid stage in the high-dose group, whereas no such changes were found in the medium- and low-dose groups. Higher serum oestradiol (P<0.05) and lower testosterone (P<0.05) levels were found in the medium- and high-dose groups. There was no significant difference in serum oestradiol and testosterone levels in the low-dose and control groups. No significant influence of OP was seen on serum FSH and LH levels in all OP-treated animals. The results from four differentially expressed proteins such as PPIA (peptidyl-prolyl cis-trans isomerase A), PEBP1 (phosphatidylethanolamine-binding protein1), TPI (triose-phosphate isomerase) and TCP-1 (T-complex protein 1) in the high-dose and control groups showed up-regulation of PPIA expression and down-regulation in PEBP1, TPI and TCP-1 expressions. These findings will contribute to clarify the mechanism of male sterility by environmental oestrogens.
Cell Biology International 11/2010; 35(4):305-9. · 1.48 Impact Factor
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Jin Zhou,
Ye Zhang,
Xinyu Wen,
Junkai Cao, Dexue Li,
Qiuxia Lin,
Haibin Wang,
Zhiqiang Liu,
Cuimi Duan,
Kuiwu Wu,
Changyong Wang
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ABSTRACT: Recently, the presence of telocytes was demonstrated in human and mammalian tissues and organs (digestive and extra-digestive organs, genitourinary organs, heart, placenta, lungs, pleura, striated muscle). Noteworthy, telocytes seem to play a significant role in the normal function and regeneration of myocardium. By cultures of telocytes in two- and three-dimensional environment we aimed to study the typical morphological features as well as functionality of telocytes, which will provide important support to understand their in vivo roles. Neonatal rat cardiomyocytes were isolated and cultured as seeding cells in vitro in two-dimensional environment. Furthermore, engineered myocardium tissue was constructed from isolated cells in three-dimensional collagen/Matrigel scaffolds. The identification of telocytes was performed by using histological and immunohistochemical methods. The results showed that typical telocytes are distributed among cardiomyocytes, connecting them by long telopodes. Telocytes have a typical fusiform cell body with two or three long moniliform telopodes, as main characteristics. The vital methylene blue staining showed the existence of telocytes in primary culture. Immunohistochemistry demonstrated that some c-kit or CD34 immuno-positive cells in engineered heart tissue had the morphology of telocytes, with a typical fusiform cell body and long moniliform telopodes. Also, a significant number of vimentin+ telocytes were present within engineered heart tissue. We suggest that the model of three-dimensional engineered heart tissue could be useful for the ongoing research on the functional relationships of telocytes with cardiomyocytes. Because the heart has the necessary potential of changing the muscle and non-muscle cells during the lifetime, telocytes might play an active role in the heart regeneration process. Moreover, telocytes might be a useful tool for cardiac tissue engineering.
Journal of Cellular and Molecular Medicine 11/2010; 14(11):2641-5. · 4.13 Impact Factor
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Shuanghong Lü,
Haibin Wang,
Wenning Lu,
Sheng Liu,
Qiuxia Lin, Dexue Li,
Cumi Duan,
Tong Hao,
Jin Zhou,
Yanmeng Wang,
Shaorong Gao,
Changyong Wang
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ABSTRACT: The transplantation of embryonic stem cells could improve cardiac function but was limited by immune rejection as well as low cell retention and survival within the ischemic tissues. The somatic cell nuclear transfer (SCNT) is practical to generate autologous histocompatible stem (nuclear-transferred embryonic stem [NTES]) cells for diseases, but NTES may be arguably unsafe for therapeutic application. The temperature-responsive chitosan hydrogel is a suitable matrix in cell transplantation. As the scaffold, chitosan hydrogel was coinjected with NTES cells into the left ventricular wall of rat infarction models. Detailed histological analysis and echocardiography were performed to determine the structure and functional consequences of transplantation. The myocardial performance in SCNT- and fertilization-derived mouse ES cell transplantation with chitosan hydrogel was also compared. The results showed that both the 24-h cell retention and 4-week graft size were significantly greater in the NTES + chitosan group than that of NTES + phosphate-buffered saline (PBS) group (p < 0.01). The NTES cells might differentiate into cardiomyocytes in vivo. The heart function improved significantly in the chitosan + NTES group (fractional shortening: 28.7% +/- 2.8%) compared with that of PBS + NTES group (fractional shortening: 25.2% +/- 2.9%) at 4 weeks after transplantation (p < 0.01). In addition, the arteriole/venule densities within the infarcted area improved significantly in the chitosan + NTES group (280 +/- 17/mm(2)) compared with that of PBS + NTES group (234 +/- 16/mm(2)) at 4 weeks after transplantation (p < 0.01). There was no difference in the myocardial performance in SCNT- and fertilization-derived mouse ES cell transplantation with chitosan hydrogel. The NTES cells with chitosan hydrogel have been proved to possess therapeutic potential to improve the function of infarcted heart. Thus the method of in situ injectable tissue engineering is promising clinically.
Tissue Engineering Part A 11/2009; 16(4):1303-15. · 4.64 Impact Factor
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ABSTRACT: B cell-specific Moloney murine leukaemia virus integration site 1 (Bmi1), known as the first functional member of PcG (Polycomb Group) family, is supposed to be a key regulator of stem cell self-proliferation. In this study, we investigated its expression in testis and its impact on spermatogonia proliferation for better understanding of its role in spermatogenesis. Results showed that Bmi1 was expressed in undifferentiated spermatogonia (A(s), A(pr) and A(al) spermatogonia). Overexpression of BMI1 could promote spermatogonia proliferation, while repression of endogenous Bmi1 by RNAi resulted in inhibition of the proliferation.
Molecular and Cellular Endocrinology 07/2008; 287(1-2):47-56. · 4.19 Impact Factor
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Shuanghong Lü,
Sheng Liu,
Wenjun He,
Cuimi Duan,
Yanmin Li,
Zhiqiang Liu,
Ye Zhang,
Tong Hao,
Yanmeng Wang, Dexue Li,
Changyong Wang,
Shaorong Gao
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ABSTRACT: Autogenic embryonic stem cells established from somatic cell nuclear transfer (SCNT) embryos have been proposed as unlimited cell sources for cell transplantation-based treatment of many genetic and degenerative diseases, which can eliminate the immune rejection that occurs after transplantation. In the present study, pluripotent nuclear transfer ES (NTES) cell lines were successfully established from different strains of mice. One NTES cell line, NT1, with capacity of germline transmission, was used to investigate in vitro differentiation into cardiomyocytes. To optimize differentiation conditions for mass production of embryoid bodies (NTEBs) from NTES cells, a slow-turning lateral vessel (STLV) rotating bioreactor was used for culturing the NTES cells to produce NTEBs compared with a conventional static cultivation method. Our results demonstrated that the NTEBs formed in STLV bioreactor were more uniform in size, and no large necrotic centers with most of the cells in NTEBs were viable. Differentiation of the NTEBs formed in both the STLV bioreactor and static culture into cardiomyocytes was induced by ascorbic acid, and the results demonstrated that STLV-produced NTEBs differentiated into cardiomyocytes more efficiently. Taken together, our results suggested that STLV bioreactor provided a more ideal culture condition, which can facilitate the formation of better quality NTEBs and differentiation into cardiomyocytes more efficiently in vitro.
Cloning and Stem Cells 07/2008; 10(3):363-70. · 2.66 Impact Factor
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Jinwu Chen,
Changyong Wang,
Shuanghong Lü,
Junzheng Wu,
Ximin Guo,
Cuimi Duan,
Lingzhi Dong,
Ying Song,
Junchuan Zhang,
Dianying Jing,
Linbo Wu,
Jiandong Ding, Dexue Li
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ABSTRACT: The purpose of this study has been to investigate the possible effects of the normal joint cavity environment on chondrocytic differentiation of bone-marrow-derived mesenchymal stem cells (MSCs). Autologous bone marrow was aspirated from the iliac crest of male sheep. MSCs were purified, expanded, and labeled with the fluorescent dye PKH26. Labeled MSCs were then grown on a three-dimensional porous scaffold of poly (L-lactic-co-glycolic acid) in vitro and implanted into the joint cavity by a surgical procedure. At 4 or 8 weeks after implantation, the implants were removed for histochemical and immunohistochemical analysis. The cells labeled with red fluorescent PKH26 in the implants expressed type II collagen and synthesized sulfated proteoglycans. However, the osteoblast-specific marker, osteocalcin, was not detected by immunohistochemistry indicating that the implanted MSCs had not differentiated into osteoblasts by being directly exposed to the normal joint cavity. To investigate the possible factors involved in chondrocytic differentiation of MSCs further, we co-cultured sheep MSCs with the main components of the normal joint cavity, viz., synovial fluid or synovial cells, in vitro. After 1 or 2 weeks of co-culture, the MSCs in both co-culture systems expressed markers of chondrogenesis. These results suggest that synovial fluid and synovium from normal joint cavity are important for the chondrocytic differentiation of adult bone-marrow-derived MSCs.
Cell and Tissue Research 04/2005; 319(3):429-38. · 3.11 Impact Factor
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ABSTRACT: Spermatogonial stem cell (SSC) transplantation is a novel technique by which testicular cells from normal, transgenic or mutant donor are introduced into the seminiferous tubules of recipient testes through microinjection. Subsequently, donor SSCs survive, migrate, anchor and proliferate in the recipient testis, furthermore, initiate spermatogenesis and even produce sperms capable of fertilization. This technique provides a new approach for the researches of spermatogenesis mechanism, regeneration of spermatogenesis in sterile individuals and reproduction of transgenic animals. This review focuses on the methodological breakthroughs and highlights the recent findings that have substantially increased understanding of SSC biology. The article provides a comprehensive overview of this technique and its multiple applications in basicscience and medicine. And the perspective direction of this field in the near future is proposed.
Progress in Natural Science 05/2004; 14(6):467-471. · 1.03 Impact Factor
