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Publications (12)

  • Ying Qi · Fengyan Zhang · Ge Song · [...] · Jian Ge
    [Show abstract] [Hide abstract] ABSTRACT: The purpose of the present study was to determine the best cholinergic neuronal differentiation method of rhesus monkey bone marrow mesenchymal stem cells (BMSCs). Four methods were used to induce differentiation, and the groups were assigned accordingly: basal inducing group (culture media, bFGF, and forskolin); SHH inducing group (SHH, inducing group); RA inducing group (RA, basal inducing group); and SHH+RA inducing group (SHH, RA, and basal inducing group). All groups displayed neuronal morphology and increased expression of nestin and neuron-specific enolase. The basal inducing group did not express synapsin, and cells from the SHH inducing group did not exhibit neuronal resting membrane potential. In contrast, results demonstrated that BMSCs from the RA and SHH+RA inducing groups exhibited neuronal resting membrane potential, and cells from the SHH+RA inducing group expressed higher levels of synapsin and acetylcholine. In conclusion, the induction of cholinergic differentiation through SHH+RA was determined to be superior to the other methods.
    Article · May 2010 · Science China. Life sciences
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    Xuerong Sun · Ruzhang Jiang · Yuehong Zhang · [...] · Jian Ge
    [Show abstract] [Hide abstract] ABSTRACT: Retinal progenitor cells (RPCs) are the most valuable seed cells in replacement therapy for neural retinal diseases. The competence of RPCs changes with retinal development. Gene expression plays a fundamental role in determining the competence. To improve the selection of the right-timing RPCs for replacement therapy, we compared the gene expression between embryonic day (E) 13.5 and E17.5 RPCs and further explored their gene expression and differentiation capacity in vitro. Timed-pregnant E13.5 and E17.5 RPCs were freshly harvested and cultured in proliferation conditions for 4 days and then in differentiation conditions for 8 days. At different time points, the expression of key genes involved in retinal development was investigated by quantitative reverse transcription-PCR or immunofluorescence. The expression of 14 key genes involved in retinal development was investigated in freshly harvested E13.5 and E17.5 RPCs. The freshly harvested E13.5 RPCs showed a high expression of retinal ganglion cell (RGC)-related genes, including Math5, Brn3b, Islet1, and Nfl, while the freshly harvested E17.5 RPCs displayed a high expression for Nrl, GFAP, and Thy1, the key genes involved in rod photoreceptor development, glial cell development, and synaptogenesis, respectively. During proliferation culture in vitro, the gene expression changed dramatically in both RPCs. After the 4 days of proliferation culture, the expression levels of most genes (11 of the 14 genes) in E13.5 RPCs came close to those in the freshly harvested E17.5 RPCs. Differentiation of RPCs in vitro was verified by the significant decrease in Nestin expression and BruU incorporation efficiency. After the 8 days of differentiation in vitro, the expression level of RGC-related genes (Math5, Brn3b, and Islet1) was still significantly higher in E13.5 RPCs than in E17.5 RPCs. In contrast, the expression level of Nrl and GFAP was significantly higher in E17.5 RPCs than in E13.5 RPCs. In morphology, the differentiated E13.5 RPCs displayed more robust process outgrowth than did the differentiated E17.5 RPCs. Immunofluorescence showed that, after the 8 days of differentiation, E13.5 RPCs contained more Brn3b- and Map2-positive cells, while E17.5 RPCs contained more GFAP-, GS-, and Rhodopsin-positive cells. The results implied that E13.5 RPCs might be a better choice for RGC replacement therapy, while E17.5 RPCs might be better for photoreceptor replacement therapy. The duration of in vitro culture should be timed, since the expression of key genes kept changing in the proliferating RPCs.
    Full-text available · Article · Dec 2009 · Molecular vision
  • Tingting Shao · Qianying Gao · Ruzhang Jiang · [...] · Jian Ge
    [Show abstract] [Hide abstract] ABSTRACT: The present study investigated dynamic alteration of low-density lipoprotein receptor and its binding and uptake of low-density lipoprotein (LDL) after exposure to transforming growth factor-beta(2) (TGF-beta(2)) in human Tenon's capsule fibroblasts. Tenon's capsule fibroblasts obtained from elective cataract surgery patients were cultured and stimulated with different concentrations (0.1-10 ng/mL) of TGF-beta(2) for 24, 48, and 72 h. The LDLr mRNA and protein levels were analyzed by relative quantification real-time RT-PCR and Western blot analysis, respectively. The binding and uptake of DiO (3,3'-dioctadecyloxacarbocyanine)-labeled LDL was assessed by confocal microscopy. Real-time RT-PCR and Western blot analyses showed similar results revealing that after exposure to TGF-beta(2), the expression of protein and mRNA of LDLr occurred in a concentration-dependent and time-dependent manner with a peak at a concentration of 1.0 ng/mL at 72 h in Tenon's capsule fibroblasts. Confocal microscopy showed that DiO-LDL binding and uptake were time-dependent, reaching saturation at approximately 6 h. This study shows that LDLrs were overexpressed in the activated Tenon's capsule fibroblasts in a concentration-dependent and time-dependent manner after exposure to TGF-beta(2). The results suggest that LDLr in the activated Tenon's capsule fibroblasts may become a novel focus as a target receptor for controlled drug delivery, particularly in anti-scarring therapy during excessive conjunctival wound healing.
    Article · Dec 2009 · Journal of ocular pharmacology and therapeutics: the official journal of the Association for Ocular Pharmacology and Therapeutics
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    Mengfei Chen · Xuerong Sun · Ruzhang Jiang · [...] · Jian Ge
    [Show abstract] [Hide abstract] ABSTRACT: Pluripotent stem cells can be induced from somatic cells by the transcription factors Oct3/4, Sox2, c-Myc and Klf4 when co-cultured with mouse embryonic fibroblast (MEF) feeder cells. To date, the role of the feeder cells in the reprogramming process remains unclear. In this study, using a comparative analysis, we demonstrated that MEF feeder cells did not accelerate reprogramming or increase the frequency of induced pluripotent stem (iPS) cell colonies. However, feeder conditions did improve the growth of primary iPS colonies and were necessary for passaging the primary colonies after reprogramming was achieved. We further developed a feeder-free culture system for supporting iPS growth and sustaining pluripotency by adding bFGF and activin A (bFA) to the medium. These data will facilitate the generation of human iPS cells without animal feeders for regenerative medicine.
    Full-text available · Article · Jul 2009 · Cell Biology International
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    Ruzhang Jiang · Bing Huang · Chenjin Jin · [...] · Jian Ge
    [Show abstract] [Hide abstract] ABSTRACT: Microinjection of adult stem cells (ASCs) into blastocysts provides a classic model for studying ASC plasticity. To explore the molecular mechanisms that govern the reprogramming of ASCs, we evaluated the experimental model through microinjection of human epidermal stem cells (hEpiSCs) into mouse blastocysts. Mouse blastocysts underwent regular embryogenesis after microinjection of allogeneic cells, confirmed by morphological observation and embryo cell counting. hEpiSCs survive and integrate into mouse embryos, by monitoring the migration of injected cells at 2, 4, 12, 16 and 24 h. In this xenogeneic system, hEpiSCs could be reprogrammed within 24 h, as evidenced by the silencing of CK15 and Integrin beta 1 gene expression, without activation of Oct4 and Nanog. Microinjection of hEpiSCs into mouse blastocysts provides an efficient model for studying the molecular mechanisms of their plasticity. Moreover, the possibility of inducing pluripotent stem cells without transgenes or viruses can be entertained.
    Full-text available · Article · Oct 2008 · Cell Biology International
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    Xuerong Sun · Jian Ge · Ruzhang Jiang · [...] · Yuehong Zhang
    [Show abstract] [Hide abstract] ABSTRACT: Key words: retinal ganglion cells, bone marrow mesenchymal stem cells, differentiation
    Full-text available · Article · Aug 2008 · Cell Research
  • Bingqian Liu · Jian Ge · Zhichong Wang · [...] · Ruzhang Jiang
    [Show abstract] [Hide abstract] ABSTRACT: limbal stem cell, ocular surface reconstruction, amniotic membrane
    Article · Aug 2008 · Cell Research
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    Ruzhang Jiang · Bing Huang · Bingqian Liu · Xuerong Sun
    [Show abstract] [Hide abstract] ABSTRACT: epidermal stem cells, chimeras, blastocysts, monkey
    Full-text available · Article · Aug 2008 · Cell Research
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    [Show abstract] [Hide abstract] ABSTRACT: Primary open-angle glaucoma (POAG) is a leading cause of irreversible blindness worldwide, and its pathogenesis is still unknown. The purpose of this study was to determine molecular changes in membrane proteins in trabecular meshwork (TM) cells from POAG patients compared to those of age-matched normal controls. Two-dimensional (2-D) gel electrophoresis profiles of membrane extracts from normal and glaucomatous TM cells were compared. The desired spots were identified after trypsin digestion and mass spectrometric analysis. Based on the results, a calcium-dependant membrane-binding protein, copine1, was further approached for a possible role in glaucomatous TM cells. The intracellular calcium concentration ([Ca(2+)]i) of TM cells was increased by incubating with calcium ionophore, A23187. Relative quantification real-time polymerase chain reaction (PCR) and western blot analysis measured copine1 expression and localization both in untreated and A23187-treated TM cells. Real-time PCR and western blot analysis confirmed that copine1 mRNA and protein expression were upregulated in glaucomatous TM cells when compared to normal ones. The cell distribution studies further showed that copine1 existed both in the membrane and cytoplasm fractions of glaucomatous TM cells but existed exclusively in cytoplasm fractions of their normal counterparts. More importantly, an influx of Ca(2+) markedly promoted the translocation of copine1 from the cytoplasm to membranes in glaucomatous TM cells. Copine1 is upregulated in plasma membranes of TM cells in individuals with POAG, which may be partly explained by its Ca(2+)-dependent translocation from the cytoplasm to the membranes. Investigation of the role and functions of copine1 in TM cells should offer new insight into the abnormal intracellular Ca(2+)-signaling pathway in glaucomatous TM and help to clarify the molecular mechanism of POAG.
    Full-text available · Article · May 2008 · Molecular vision
  • [Show abstract] [Hide abstract] ABSTRACT: Retinoblastoma (RB) is the most common malignant tumor of the retina in human children. Although it has been hypothesized for a long time that RB derives from multipotent retinal stem cells (RSCs) or retinoblasts, the direct evidence that the presence of tumorigenic RSCs in RB tumors is still lacking. Some studies indicate that malignant tumors contain tumor stem cells similar to their normal tissue stem cell counterparts. With in vitro culture and differentiation method we demonstrate that tumorigenic retinal stem-like cells (RSLCs) indeed exist in RB lesions and that RB tumor-derived cultures encompass undifferentiated cells capable of extensive proliferation as clonal nonadherent neurospheres and can differentiate into different retinal cells in vitro. Interestingly, cultured cells expressed retinal development related genes including nestin, CD133, pax6, chx10 and Rx, and overexpressed Bmi-1, a gene required for self-renewal and proliferation of stem cells. Significantly, when these cultured cells were intraocularly transplanted into SCID mice, they gave rise to new tumors with histomorphological features and immunophenotypes similar to their parental primary RBs. The results show that RBs contain tumorigenic RSLCs that contribute to tumorigenesis. This study provides a new insight to investigate the histogenesis of RBs and establishes a model for other RB research.
    Article · Nov 2007 · International Journal of Cancer
  • [Show abstract] [Hide abstract] ABSTRACT: Bioengineered corneas are substitutes for human donor tissue that are designed to treat severe disease affecting ocular surfaces. However, a shortage of candidate seed cells for bioengineering corneas is still a problem. Bone-marrow mesenchymal stem cells (MSCs) are capable of multilineage differentiation. Therefore, we determined whether MSCs differentiate into corneal epithelial cells (ECs). We applied three exoteric-microenvironmental systems to induce MSCs to become ECs. Induced MSC were identified by means of morphologic examination, immunocytochemical analysis, and flow cytometry. MSCs grown in one microenvironment had characteristics similar to those of corneal epithelial progenitors. Induced MSCs expressed markers for EC, including integrin β1, Cx43, Pax6, and P63. MSCs were successfully induced to become corneal epithelial progenitors. Therefore, the use of MSCs may hold substantial promise for reconstructing the ocular surface after corneal injury.
    Article · Aug 2007 · Chinese Science Bulletin
  • [Show abstract] [Hide abstract] ABSTRACT: To evaluate short-term effects of single photodynamic therapy (PDT) for age-related macular degeneration (AMD) accompanied with choroidal neovascularization (CNV). We analyzed retrospectively the effects of single PDT for 20 patients (20 eyes) with CNV caused by AMD. Corrected visual acuity, fluorescein angiography (FA) and optic coherence tomography (OCT) were examined before and after PDT. All patients were followed up at least 3 months. At the end of 3-month follow-up, 5 eyes had vision progress, 15 eyes had vision stable and no eye had vision deterioration. Fluorescein angiography one week post PDT showed cessation of fluorescein leakage in 8 eyes with predominant classic CNV, and reduction of fluorescein leakage in 12 eyes with minimal classic CNV or occult CNV without classic component. At the 3-month following PDT fluorescein angiography showed fluorescein leakage reappeared in 4 of 8 eyes with predominant classic CNV. Among 12 eyes with minimal classic CNV or occult CNV without classic component, 9 eyes showed decreased or unchanged fluorescein leakage, 3 eyes had a progression of fluorescein leakage. Optic coherence tomography showed obvious recovery of serous sensory retinal detachment after PDT. PDT may occlude or inhibit CNV caused by AMD in short-term. No obvious side effects were noticed.
    Article · Oct 2004 · Yan ke xue bao = Eye science / "Yan ke xue bao" bian ji bu