Published literature

Stem Cell (13) Products

  Human OneArray  
 Scientific Reports. 2015, 5:10106. doi: 10.1038/srep10106.
 Characterization of a Self-renewing and Multi-potent Cell Population Isolated from Human Minor Salivary Glands 
 Lin Lu, Yan Li, Ming-juan Du, Chen Zhang, Xiang-yu Zhang, Hai-zhou Tong, Lei Liu, Ting-lu Han, Wan-di Li, Li Yan, Ning-bei Yin, Hai-dong Li, Zhen-min Zhao
  Abstract
Adult stem cells play an important role in maintaining tissue homeostasis. Although these cells are found in many tissues, the presence of stem cells in the human minor salivary glands is not well explored. Using the explant culture method, we isolated a population of cells with self-renewal and differentiation capacities harboring that reside in the human minor salivary glands, called human minor salivary gland mesenchymal stem cells (hMSGMSCs). These cells show embryonic stem cell and mesenchymal stem cell phenotypes. Our results demonstrate that hMSGMSCs have the potential to undergo mesodermal, ectodermal and endodermal differentiation in conditioned culture systems in vitro. Furthermore, in vivo transplantation of hMSGMSCs into SCID mice after partial hepatectomy shows that hMSGMSCs are able to survive and engraft, characterized by the survival of labeled cells and the expression of the hepatocyte markers AFP and KRT18. These data demonstrate the existence of hMSGMSCs and suggest their potential in cell therapy and regenerative medicine.
   

  Human OneArray  
 Northeast Bioengineering Conference (NEBEC). 2014 April 25-27.
 FGF2 and oxygen: Regulators of intergrin alpha-11 and extracellular matrix molecules
 
 
 Alexandra Grella, Denis Kole, Tanja Dominko
  Abstract
Recently, derivation and maintenance of pluripotent stem cells has been focused on environmental cues, with emphasis on the role of extracellular matrix (ECM) and adhesion molecules (AM). We have developed a novel approach that allows for induction of stem cell gene expression in human dermal fibroblasts (hDF) without the use of transgenes. By culturing cells in low oxygen (5% O2) with addition of exogenous FGF2 we have shown that hDF in defined culture conditions express stem cell genes and show translation and nuclear translocation of stem cell transcription factors. We have demonstrated that this shift is coupled with an FGF2-dependent down-regulation of the majority of AM and ECM targets; specifically induction of a significant down-regulation of integrin alpha 11 (Itga11) transcript and results in Itga11 loss from focal adhesions. Investigation of the mechanism by which FGF2 may be involved in regulation of Itga11 is being investigated by studying the molecular pathway downstream of FGF2 ligand that may be involved in the loss of Itga11 and associated collagen I attachment. Dissecting the molecular mechanisms involved in regulation through modulation of extracellular environment and its effect on plasticity may provide insight into the acquisition into the mechanisms involved in reprogramming of differentiated cells.
   

  Human OneArray  
 Medical Journal of Chinese People's Liberation Army. 2014 Oct 1. doi: 10.11855/j.issn.0577-7402.2014.10.08.
 Identification of the difference in gene expression between glioma stem cells and neural stem cells by oligonucleotide microarray
 
 
 Shuang LIU, Feng YIN, Jian-ning ZHANG, Ming-ming ZHAO, Chun-hui ZHOU, Shu-wei WANG, Xin-ru GUO
  Abstract
ObjectiveTo identify the differential expressed genes of human glioma stem cells (GSCs) and human neural stem cells (NSCs) by gene chip technology. MethodsHuman HOA5.1 OneArray microarray (including 29 186 genes) was adopted and hybridized with probes which were prepared from the total RNAs of GSCs and NSCs. Differential expressed genes between the GSCs and NSCs were assayed after scanning oligonucleotide microarray with ScanArray 4000, and some of these genes such as DCX, PTGS2, SCGN, GAD2, OTX2, PEG10 and NRXN3 were verified by real-time-Q-PCR method. ResultsCompared with the genes in normal NSCs, 1372 down-regulated and 1501 up-regulated genes in GSCs were revealed by means of microarray, and these genes were associated with axon guidance, cell cycle, cell adhesion, immune-inflammatory responses and cancer-related signal pathways. The results of qRT-PCR were consistent with that of microarray. ConclusionsMultiple genes play important roles in development of glioma. This study may provide new clues for the targeted therapy of malignant glioma.
   

  Human miRNA OneArray  
 Journal of Cellular Biochemistry. 2014 Feb 12. doi: 10.1002/jcb.24786.
 Regulatory Roles of miRNA in the Human Neural Stem Cell Transformation to Glioma Stem Cells
 
 
 Shuang Liu, Jianning Zhang, Max S. Wicha, Alfred E. Chang, Wenhong Fan, Ling Chen, Ming Fan, Qiao Li, Feng Yin
  Abstract
To investigate the expressional alternation of microRNAs (miRNA) during the malignant transformation and development of human glioma, we measuredmiRNA expression profile as well as mRNA expression profile in normal human neural stem cells (hNSCs) and human glioma stem cells (hGSCs). We found 116 miRNA up-regulated and 62 miRNA down-regulated in GSCs. On the other hand, we identified 1,372 mRNA down-regulated, and 1,501 mRNA up-regulated in GSCs compared to those in NSCs. We then analyzed the pathways and the predicted target genes of the miRNAs which differ significantly in expression between GSCs and NSCs using the statistical enrichment methods. These target mRNAs are involved in many cancer-related signaling pathways, such as cell cycle, axon guidance, glioma development, adhesion junction, MAPK and Wnt signaling. Furthermore, we obtained the differently expressed miRNA-target relationships according to the 庛 value which is used to calculate the regulation extent of miRNA-target and using the databases of miRanda, Targetscans and Pictar. Among the top 10 miRNA-target relationships, hsa-miR-198 and its potential targeted gene DCX and NNAT were selected for validation, and NNAT was found to be the direct target of miR-198. Finally, the functional roles of miR-155-5p and miR-124-3p whose expressions altered significantly between GSCs and NSCs were addressed. Our results provide new clues for the potential mechanisms involved in the origin and development of glioma. Clinically, the altered miRNAs may serve as potential targets and diagnostic tools for novel therapeutic strategies of glioblastoma.
   

  Human miRNA OneArray  
 Nature Cell Biology. 2014 Feb 23;16(3):268-280. doi: 10.1038/ncb2910.
 MicroRNA-146a directs the symmetric division of Snail-dominant colorectal cancer stem cells
 
 
 Wei-Lun Hwang, Jeng-Kae Jiang, Shung-Haur Yang, Tse-Shun Huang, Hsin-Yi Lan, Hao-Wei Teng, Chih-Yung Yang, Ya-Ping Tsai, Chi-Hung Lin, Hsei-WeiWang, Muh-Hwa Yang
  Abstract
Asymmetrical cell division (ACD) maintains the proper number of stem cells to ensure self-renewal. In cancer cells, the deregulation of ACD disrupts the homeostasis of the stem cell pool and promotes tumour growth. However, this mechanism is unclear. Here, we show a reduction of ACD in spheroid-derived colorectal cancer stem cells (CRCSCs) compared with differentiated cancer cells. The epithelial-mesenchymal transition (EMT) inducer Snail is responsible for the ACD-to-symmetrical cell division (SCD) switch in CRCSCs. Mechanistically, Snail induces the expression of microRNA-146a (miR-146a) through the 帣-catenin-TCF4 complex. miR-146a targets Numb to stabilize 帣-catenin, which forms a feedback circuit to maintain Wnt activity and directs SCD. Interference with the Snail-miR-146a-帣-catenin loop by inhibiting the MEK or Wnt activity reduces the symmetrical division of CRCSCs and attenuates tumorigenicity. In colorectal cancer patients, the SnailHighNumbLow profile is correlated with cetuximab resistance and a poorer prognosis. This study elucidates a unique mechanism of EMT-induced CRCSC expansion.
   

  Mouse OneArray  
 Science. 2013, 341(6146): 651-654. doi: 10.1126/science.1239278.
 Pluripotent Stem Cells Induced from Mouse Somatic Cells by Small-Molecule Compounds
 
 
 Pingping Hou, Yanqin Li, Xu Zhang, Chun Liu, Jingyang Guan, Honggang Li, Ting Zhao, Junqing Ye, Weifeng Yang, Kang Liu, Jian Ge, Jun Xu, Qiang Zhang, Yang Zhao, Hongkui Deng
  Abstract
Pluripotent stem cells can be induced from somatic cells, providing an unlimited cell resource, with potential for studying disease and use in regenerative medicine. However, genetic manipulation and technically challenging strategies such as nuclear transfer used in reprogramming limit their clinical applications. Here, we show that pluripotent stem cells can be generated from mouse somatic cells at a frequency up to 0.2% using a combination of seven small-molecule compounds. The chemically induced pluripotent stem cells (CiPSCs) resemble embryonic stem cells (ESCs) in terms of their gene expression profiles, epigenetic status, and potential for differentiation and germline transmission. By using small molecules, exogenous master genes are dispensable for cell fate reprogramming. This chemical reprogramming strategy has potential use in generating functional desirable cell types for clinical applications.
   

  Mouse OneArray  
 Cell. 2013, 153(5): 963-975. doi: 10.1016/j.cell.2013.05.001.
 Induction of Pluripotency in Mouse Somatic Cells with Lineage Specifiers
 
 
 Jian Shu, Chen Wu, Yetao Wu, Zhiyuan Li, Sida Shao, Wenhui Zhao, Xing Tang, Huan Yang, Lijun Shen, Xiaohan Zuo, Weifeng Yang, Yan Shi, Xiaochun Chi, Hongquan Zhang, Ge Gao, Youmin Shu, Kehu Yuan, Weiwu He, Yang Zhao, Chao Tang, Hongkui Deng
  Abstract
The reprogramming factors that induce pluripotency have been identified primarily from embryonic stem cell (ESC)-enriched, pluripotency-associated factors. Here, we report that, during mouse somatic cell reprogramming, pluripotency can be induced with lineage specifiers that are pluripotency rivals to suppress ESC identity, most of which are not enriched in ESCs. We found that OCT4 and SOX2, the core regulators of pluripotency, can be replaced by lineage specifiers that are involved in mesendodermal (ME) specification and in ectodermal (ECT) specification, respectively. OCT4 and its substitutes attenuated the elevated expression of a group of ECT genes, whereas SOX2 and its substitutes curtailed a group of ME genes during reprogramming. Surprisingly, the two counteracting lineage specifiers can synergistically induce pluripotency in the absence of both OCT4 and SOX2. Our study suggests a seesaw model in which a balance that is established using pluripotency factors and/or counteracting lineage specifiers can facilitate reprogramming.
   

  Human miRNA OneArray  
 Oncology Reports. 2012, 28(6):2115-24. doi: 10.3892/or.2012.2054.
 miRNA expression profile of colon cancer stem cells compared to non-stem cells using the SW1116 cell line
 
 
 ZONGYOU CHEN, YANTIAN FANG, JIANBIN XIANG, XIAODONG GU, ZHENGYANG LI, FENG TANG, ZHONGWEN ZHOU
  Abstract
Colorectal cancer (CRC) is one of the major causes of cancer-related mortality worldwide. Recent studies revealed that there is a relationship between CRC occurrence and microRNA (miRNA) function. Stem cells are a type of cells that have the ability to self-renew and to proliferate extensively while maintaining the undifferentiated state. Cancer stem cells (CSCs) are closely linked to tumor recurrence and metastasis. To this end, we evaluated themiRNA expression differences between colon CSCs and non-stem cells using the SW1116 cell line, to determine the relationship between tumorstem cells and tumor biological behavior. We isolated populations of colon CSCs with the CD133+/CD44+ and CD133-/CD44- surface phenotype from a human SW1116 colon adenocarcinoma cell line using flow cytometry. The expression of miRNA and mRNA of both sets of cells was examined withmiRNA and mRNA arrays. Bioinformatic methods were used to analyze microarray results. We completed gene ontology analysis, pathway analysis,miRNA target gene prediction with databases. We identified a colon stem cell miRNA expression profile comprising 31 upregulated and 31 downregulated miRNAs, such as miR29a, miR29b, miR449b and miR4524. Some of these differentially expressed miRNAs may be involved in the regulation of stem cell differentiation. Gene ontology and pathway analyses showed that the differences are closely related to the function of the cellcycle, cell differentiation, signaling pathway, cytoskeletal proteins and cell-matrix adhesion in colon cancer stem cells. We found that miRNAs play an important role in regulating the expression of colon CSC characteristics. By regulating the expression of CSC signaling pathways, cytoskeleton and membrane proteins, miRNAs give tumor stem cells the macrobiological behavior of recurrence and metastasis. This study provides a new perspective on CRC metastasis and recurrence.
   

  Experimental Accessories  
 Journal of Virology. 2012, 86(15):8041-9. doi: 10.1128/JVI.00808-12.
 Defective Antiviral Responses of Induced Pluripotent Stem Cells to Baculoviral Vector Transduction
 
 
 Chen GY, Hwang SM, Su HJ, Kuo CY, Luo WY, Lo KW, Huang CC, Chen CL, Yu SH, Yu-Chen Hu
  Abstract
Genetic engineering of induced pluripotent stem cells (iPSCs) is important for their clinical applications, and baculovirus (BV) holds promise as a gene delivery vector. To explore the feasibility of using BV for iPSCs transduction, in this study we first examined how iPSCs responded to BV. We determined that BV transduced iPSCs efficiently, without inducing appreciable negative effects on cell proliferation, apoptosis, pluripotency, and differentiation. BV transduction slightly perturbed the transcription of 12 genes involved in the Toll-like receptor (TLR) signaling pathway, but at the protein level BV elicited no well-known cytokines (e.g., interleukin-6 [IL-6], tumor necrosis factor alpha [TNF-], and beta interferon [IFN-]) except for IP-10. Molecular analyses revealed that iPSCs expressed no TLR1, -6, -8, or -9 and expressed merely low levels of TLR2, -3, and -4. In spite of evident expression of such RNA/DNA sensors as RIG-I and AIM2, iPSCs barely expressed MDA5 and DAI (DNA-dependent activator of IFN regulatory factor [IRF]). Importantly, BV transduction of iPSCs stimulated none of the aforementioned sensors or their downstream signaling mediators (IRF3 and NF-B). These data together confirmed that iPSCs responded poorly to BV due to the impaired sensing and signaling system, thereby justifying the transduction of iPSCs with the baculoviral vector.
   

  Mouse OneArray  
 Transgenic Res. 2011, 20(5):1073-86. doi: 10.1007/s11248-010-9478-2.
 Epithelial cell-targeted transgene expression enables isolation of cyan fluorescent protein (CFP)-expressing prostate stem/progenitor cells
 
 
 Weidan Peng, Yunhua Bao, Janet A. Sawicki
  Abstract
To establish a method for efficient and relatively easy isolation of a cell population containing epithelial prostate stem cells, we developed two transgenic mouse models, K5/CFP and K18/RFP. In these models, promoters of the cytokeratin 5 (Krt5) and the cytokeratin 18 (Krt18) genes regulate cyan and red fluorescent proteins (CFP and RFP), respectively. CFP and RFP reporter protein fluorescence allows for visualization of K5(+) and K18(+) epithelial cells within the cellular spatial context of the prostate gland and for their direct isolation by FACS. Using these models, it is possible to test directly the stem cell properties of prostate epithelial cell populations that are positively selected based on expression of cytoplasmic proteins, K5 and K18. After validating appropriate expression of the K5/CFP and K18/RFP transgenes in the developing and adult prostate, we demonstrate that a subset of CFP-expressing prostate cells exhibits stem cell proliferation potential and differentiation capabilities. Then, using prostate cells sorted from double transgenic mice (K5/CFP + K18/RFP), we compare RNA microarrays of sorted K5(+)K18(+) basal and K5(-)K18(+) luminal epithelial cells, and identify genes that are differentially expressed. Several genes that are over-expressed in K5(+) cells have previously been identified as potential stem cell markers. These results suggest that FACS isolation of prostate cells from these mice based on combining reporter gene fluorescence with expression of potential stem cell surface marker proteins will yield populations of cells enriched for stem cells to a degree that has not been attained by using cell surface markers alone.
   

  Mouse OneArray  
 Cell Research. 2011, 21(1):196-204. doi: 10.1038/cr.2010.142.
 Generation of iPSCs from mouse fibroblasts with a single gene, Oct4, and small molecules.
 
 
 Yanqin Li , Qiang Zhang , Xiaolei Yin , Weifeng Yang , Yuanyuan Du , Pingping Hou , Jian Ge , Chun Liu, Weiqi Zhang, Xu Zhang1,, Yetao Wu1,, Honggang Li, Kang Liu, Chen Wu1,, Zhihua Song, Yang Zhao, Yan Shi, Hongkui Deng.
  Abstract
The introduction of four transcription factors Oct4, Klf4, Sox2 and c-Myc by viral transduction can induce reprogramming of somatic cells into induced pluripotent stem cells (iPSCs), but the use of iPSCs is hindered by the use of viral delivery systems. Chemical-induced reprogramming offers a novel approach to generating iPSCs without any viral vector-based genetic modification. Previous reports showed that several small molecules could replace some of the reprogramming factors although at least two transcription factors, Oct4 and Klf4, are still required to generate iPSCs from mouse embryonic fibroblasts. Here, we identify a specific chemical combination, which is sufficient to permit reprogramming from mouse embryonic and adult fibroblasts in the presence of a single transcription factor, Oct4, within 20 days, replacing Sox2, Klf4 and c-Myc. The iPSCs generated using this treatment resembled mouse embryonic stem cells in terms of global gene expression profile, epigenetic status and pluripotency both in vitro and in vivo. We also found that 8 days of Oct4 induction was sufficient to enable Oct4-induced reprogramming in the presence of the small molecules, which suggests that reprogramming was initiated within the first 8 days and was independent of continuous exogenous Oct4 expression. These discoveries will aid in the future generation of iPSCs without genetic modification, as well as elucidating the molecular mechanisms that underlie the reprogramming process.
   

  Human OneArray  
 JOURNAL OF VIROLOGY. 2009, 83(20):10548-56. doi: 10.1128/JVI.01250-09.
 Baculovirus Transduction of Mesenchymal Stem Cells Triggers the Toll-Like Receptor 3 Pathway.
 
 
 Guan-Yu Chen, Hsiao-Chiao Shiah, Hung-Ju Su, Chi-Yuan Chen, Yung-Jen Chuang, Wen-Hsin Lo, Jie-Len Huang, Ching-Kuang Chuang, Shiaw-Min Hwang, Yu-Chen Hu
  Abstract
Human mesenchymal stem cells (hMSCs) can be genetically modified with viral vectors and hold promise as a cell source for regenerative medicine, yet how hMSCs respond to viral vector transduction remains poorly understood, leaving the safety concerns unaddressed. Here, we explored the responses of hMSCs against an emerging DNA viral vector, baculovirus (BV), and discovered that BV transduction perturbed the transcription of 816 genes associated with five signaling pathways. Surprisingly, Toll-like receptor-3 (TLR3), a receptor that generally recognizes double-stranded RNA, was apparently upregulated by BV transduction, as confirmed by microarray, PCR array, flow cytometry, and confocal microscopy. Cytokine array data showed that BV transduction triggered robust secretion of interleukin-6 (IL-6) and IL-8 but not of other inflammatory cytokines and beta interferon (IFN-beta). BV transduction activated the signaling molecules (e.g., Toll/interleukin-1 receptor domain-containing adaptor-inducing IFN-beta, NF-kappaB, and IFN regulatory factor 3) downstream of TLR3, while silencing the TLR3 gene with small interfering RNA considerably abolished cytokine expression and promoted cell migration. These data demonstrate, for the first time, that a DNA viral vector can activate the TLR3 pathway in hMSCs and lead to a cytokine expression profile distinct from that in immune cells. These findings underscore the importance of evaluating whether the TLR3 signaling cascade plays roles in the immune response provoked by other DNA vectors (e.g., adenovirus). Nonetheless, BV transduction barely disturbed surface marker expression and induced only transient and mild cytokine responses, thereby easing the safety concerns of using BV for hMSCs engineering.
   

  Mouse OneArray  
 Cell Stem Cell. 2008, 3(5):475-9. doi: 10.1016/j.stem.2008.10.002.
 Two Supporting Factors Greatly Improve the Efficiency of Human iPSC Generation.
 
 
 hao Y, Yin X, Qin H, Zhu F, Liu H, Yang W, Zhang Q, Xiang C, Hou P, Song Z, Liu Y, Yong J, Zhang P, Cai J, Liu M, Li H, Li Y, Qu X, Cui K, Zhang W, Xiang T, Wu Y, Zhao Y, Liu C, Yu C, Yuan K, Lou J, Ding M, Deng H.
  Abstract
Human ?broblasts can be induced into pluripotent stem cells (iPSCs), but the reprogramming ef?ciency is quite low. Here, we screened a panel of candidate factors in the presence of OCT4, SOX2, KLF4, and c-MYC in an effort to improve the reprogramming ef?ciency from human adult ?broblasts. We found that p53 siRNA and UTF1 enhanced the ef?ciency of iPSC generation up to 100-fold, even when the oncogene c-MYC was removed from the combinations.