ABSTRACT
Pulmonary fibrosis (PF) is a progressive and irreversible condition with various causes, and no effective treatment has been found to rescue fibrotic lungs. Successful recovery from PF requires inhibiting inflammation, promoting collagen degradation and stimulating alveolar regeneration. Human umbilical mesenchymal stem cells (HUMSCs) not only regulate immune responses but also synthesize and release hyaluronan to improve lung regeneration. This study investigated the feasibility of HUMSC engraftment into rats with bleomycin (BLM)-induced PF to explore HUMSC therapeutic effects/outcomes. Methods: A unique BLM-induced left-lung-dominated PF animal model was established. Rats were transplanted with low-dose (5×106) or high-dose (2.5×107) HUMSCs on Day 21 after BLM injection. Combinations in co-culture of pulmonary macrophages, fibroblasts, HUMSCs treated with BLM and the same conditions on alveolar epithelia versus HUMSCs were evaluated. Results: Rats with high-dose HUMSC engraftment displayed significant recovery, including improved blood oxygen saturation levels and respiratory rates. High-dose HUMSC transplantation reversed alveolar injury, reduced cell infiltration and ameliorated collagen deposition. One month posttransplantation, HUMSCs in the rats' lungs remained viable and secreted cytokines without differentiating into alveolar or vascular epithelial cells. Moreover, HUMSCs decreased epithelial-mesenchymal transition in pulmonary inflammation, enhanced macrophage matrix-metallopeptidase-9 (MMP-9) expression for collagen degradation, and promoted toll-like receptor-4 (TLR-4) expression in the lung for alveolar regeneration. In coculture studies, HUMSCs elevated the MMP-9 level in pulmonary macrophages, released hyaluronan into the medium and stimulated the TLR-4 quantity in the alveolar epithelium. Principal Conclusions: Transplanted HUMSCs exhibit long-term viability in rat lungs and can effectively reverse rat PF.
Subject(s)
Mesenchymal Stem Cell Transplantation/methods , Mesenchymal Stem Cells/metabolism , Pulmonary Fibrosis/therapy , Wharton Jelly/cytology , Animals , Bleomycin/toxicity , Cell Differentiation , Cytokines/metabolism , Disease Models, Animal , Heterografts , Humans , Male , Matrix Metalloproteinase 9/metabolism , Mesenchymal Stem Cells/cytology , Pulmonary Fibrosis/chemically induced , Pulmonary Fibrosis/metabolism , Pulmonary Fibrosis/pathology , Pulmonary Gas Exchange , Rats, Sprague-Dawley , Respiratory Function Tests , Toll-Like Receptor 4/metabolism , Transplantation, Heterologous , Umbilical Cord/cytologyABSTRACT
The success rate in previous attempts at transforming human umbilical mesenchymal stem cells (HUMSCs) isolated from Wharton's jelly of the umbilical cord into dopaminergic cells was a mere 12.7%. The present study was therefore initiated to establish a more effective procedure for better yield of dopaminergic cells in such transformation for more effective HUMSC-based therapy for parkinsonism. To examine, in vitro, the effects of enhanced Nurr1 expression in HUMSCs on their differentiation, cells were processed through the three-stage differentiation protocol. The capacity of such cells to synthesize and release dopamine was measured by HPLC. The therapeutic effects of Nurr1-overexppressed HUMSCs were examined in 6-hydroxydopamine-lesioned rats by quantification of rotations in response to amphetamine. Enhanced Nurr1 expression in HUMSCs promoted the transformation into dopaminergic cells in vitro through stepwise culturing in sonic hedgehog, fibroblast growth factor-8, and neuron-conditioned medium. The success rate was about 71%, as determined by immunostaining for tyrosine hydroxylase and around 94 nM dopamine synthesis (intracellular and released into the culture medium), as measured by HPLC. Additionally, transplantation of such cells into the striatum of hemiparkinsonian rats resulted in improvement of their behavioral deficits, as indicated by amphetamine-evoked rotation scores. Viability of the transplanted cells lasted for at least 3 months as verified by positive staining for tyrosine hydroxylase. Nurr1, FGF8, Shh, and NCM can synergistically enhance the differentiation of HUMSCs into dopaminergic cells and may pave the way for HUMSC-based treatments for Parkinson's disease.
Subject(s)
Cell Differentiation , Dopaminergic Neurons/transplantation , Mesenchymal Stem Cells/cytology , Parkinsonian Disorders/therapy , Wharton Jelly/cytology , Animals , Cell Culture Techniques , Disease Models, Animal , Dopamine/biosynthesis , Humans , Male , Nuclear Receptor Subfamily 4, Group A, Member 2/genetics , Parkinsonian Disorders/physiopathology , Rats , Rats, Sprague-Dawley , Tyrosine 3-Monooxygenase/metabolism , Umbilical Cord/cytologyABSTRACT
The aim of this experiment is to identify related genes for human umbilical mesenchymal stem cells transformation into nervous cells. After the human umbilical mesenchymal stem cells were treated with neuronal conditioned medium (NCM) for 9 days, the gene expression groups are compared to those only treated with DMEM. The related genes for cell cycles, the human umbilical mesenchymal stem cells treated with DMEM increases the amount of cells that remain in the G2/M phase and S phase, including CAV1, EBF, NRG1, CDH13, MLH1. After treatment, the human umbilical cord mesenchymal stem cells with NCM for 9 days, gene expression related to the G0/G1 phase are also increased, including MYC, CSF3, PETN. Gene expressions related to neural regeneration and neural stem cells also increase significantly, such as CXCL1, BMP2, NRCAM, FGF2, SPG7. This study thereby provides a foundation for a more detailed understanding of HUMSCs neuronal differentiation.
Subject(s)
Gene Expression Regulation, Developmental , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/physiology , Neurogenesis/genetics , Neurons/cytology , Oligonucleotide Array Sequence Analysis , Animals , Cell Culture Techniques , Cell Differentiation/genetics , Culture Media, Conditioned/pharmacology , Cytokines/genetics , Fetal Blood/cytology , Gene Expression Profiling , Humans , Rats , Rats, Sprague-DawleyABSTRACT
We tested the herbal extract 2,3,5,6-tetramethylpyrazine (TMP) for possible therapeutic efficacy against a glioma cell line and against gliomas transplanted into rat brains. In the cultured glioma cells, 50 muM TMP significantly inhibited glutamate-induced increase in intracellular calcium. Significant cell damage (30%) and proliferation suppression (10%), however, occurred only at higher concentrations (200-400 microM). Gliomaneuronal co-culturing resulted in significant neuronal damage and higher proliferation of the glioma cells (140%) compared with single cultures. Low concentrations of TMP (< or =200 microM) attenuated the neuronal damage, suppressed glioma migration, and decreased glioma proliferation in the neuronal-glioma co-culture. Gliomas transplanted into the frontal cortical area exhibited high proliferation, with untreated rats dying 10-23 days later. TMP treatment inhibited tumor growth and significantly extended survival time. The results indicate that TMP can suppress glioma activity, including growth, and protect neurons against glioma-induced excitotoxicity, suggesting that TMP may have therapeutic potential in the treatment of malignant gliomas.
