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Guangchuan Wang, Guang Yu, Dongmei Wang, Shengnan Guo, Fengping Shan
Natural killer (NK) cells are innate lymphocytes that aid in the protection of the host from infectious diseases and cancer. In vitro studies of NK cells have provided a foundation for developing clinical adoptive NK-cell transferred immunotherapy against human tumors. To elucidate the functions and mechanisms of NK cell populations, it is important to develop an optimal, highly reproducible and reliable isolation method. The present comparative study was performed with four different NK cell isolation kits of magnetic bead labeling made by Miltenyi and Stemcell companies, including positive selection kits [cluster of differentiation (CD)-49b, using the monoclonal antibody DX5) MicroBeads] and negative selection kits...
May 2017: Experimental and Therapeutic Medicine
Kieran O'Neill, Mark Hills, Mike Gottlieb, Matthew Borkowski, Aly Karsan, Peter M Lansdorp
Summary: Massively parallel sequencing is now widely used, but data interpretation is only as good as the reference assembly to which it is aligned. While the number of reference assemblies has rapidly expanded, most of these remain at intermediate stages of completion, either as scaffold builds, or as chromosome builds (consisting of correctly ordered, but not necessarily correctly oriented scaffolds separated by gaps). Completion of de novo assemblies remains difficult, as regions that are repetitive or hard to sequence prevent the accumulation of larger scaffolds, and create errors such as misorientations and mislocalizations...
September 1, 2017: Bioinformatics
Maren Bredemeier, Sabine Kasimir-Bauer, Hans-Christian Kolberg, Thomas Herold, Sarah Synoracki, Siegfried Hauch, Philippos Edimiris, Agnes Bankfalvi, Mitra Tewes, Rainer Kimmig, Bahriye Aktas
The aim of the present study was to compare the phosphatidylinositol 3-kinase (PI3KCA)-AKT serine/threonine kinase (AKT) pathway in circulating tumor cells (CTCs) and corresponding cancerous tissues. Stemness‑like circulating tumor cells (slCTCs) and CTCs in epithelial-mesenchymal transition (EMT) have been implicated as the active source of metastatic spread in breast cancer (BC). In this regard, the PI3KCA‑AKT signaling pathway was demonstrated to be implicated in and to be frequently mutated in BC. The present study compared this pathway in slCTCs/CTCs in EMT and the corresponding tumor tissues of 90 metastatic BC patients (pts)...
May 2017: Molecular Medicine Reports
Sung-Hoon Han, Sehwan Shim, Min-Jung Kim, Hye-Yun Shin, Won-Suk Jang, Sun-Joo Lee, Young-Woo Jin, Seung-Sook Lee, Seung Bum Lee, Sunhoo Park
AIM: To investigate a suitable long-term culture system and optimal cryopreservation of intestinal organoid to improve organoid-based therapy by acquiring large numbers of cells. METHODS: Crypts were isolated from jejunum of C57BL/6 mouse. Two hundred crypts were cultured in organoid medium with either epidermal growth factor/Noggin/R-spondin1 (ENR) or ENR/CHIR99021/VPA (ENR-CV). For subculture, organoids cultured on day 7 were passaged using enzyme-free cell dissociation buffer (STEMCELL Technologies)...
February 14, 2017: World Journal of Gastroenterology: WJG
Samuel E Marsh, Stephen T Yeung, Maria Torres, Lydia Lau, Joy L Davis, Edwin S Monuki, Wayne W Poon, Mathew Blurton-Jones
Transplantation of neural stem cells (NSCs) can improve cognition in animal models of Alzheimer's disease (AD). However, AD is a protracted disorder, and prior studies have examined only short-term effects. We therefore used an immune-deficient model of AD (Rag-5xfAD mice) to examine long-term transplantation of human NSCs (StemCells Inc.; HuCNS-SCs). Five months after transplantation, HuCNS-SCs had engrafted and migrated throughout the hippocampus and exhibited no differences in survival or migration in response to β-amyloid pathology...
February 14, 2017: Stem Cell Reports
Qing Tian, Shilei Wu, Zhipeng Dai, Jingjing Yang, Jin Zheng, Qixin Zheng, Yong Liu
BACKGROUND: Iron overload is recognized as a new pathogenfor osteoporosis. Various studies demonstrated that iron overload could induce apoptosis in osteoblasts and osteoporosis in vivo. However, the exact molecular mechanisms involved in the iron overload-mediated induction of apoptosis in osteoblasts has not been explored. PURPOSE: In this study, we attempted to determine whether the mitochondrial apoptotic pathway is involved in iron-induced osteoblastic cell death and to investigate the beneficial effect of N-acetyl-cysteine (NAC) in iron-induced cytotoxicity...
2016: PeerJ
V Rylova, L B Buravkova, B D Zhivotovky
OBJECTIVE: To evaluate the damaging effects of cisplatin on MMSCs from adipose tissue in a phase of active proliferation and the state of the monolayer, this was exposed at standard (20%) and reduced to 1% and 5% level of oxygen. METHODS: The effect of cisplatin was detected on monolayer cultures in the active growth phase after 2 passages. Profile surface markers of MMSC was determined by flow cytometry. MMSCs viability after incubation with cisplatin was detected by the number of apoptotic and necrotic cells using ANNEXIN V-FITC--PI Kit (Immunotech, France)...
2016: Vestnik Rossiĭskoĭ Akademii Meditsinskikh Nauk
Akemi Ota, Kazuaki Matsumura, Jun-Jae Lee, Shoichiro Sumi, Soung-Hyu Hyon
Safe and stable cryopreservation is critical for research involving human embryonic stem cells (hESCs). Dimethyl sulfoxide (DMSO) is a popular cryoprotective agent; however, its cytotoxicity cannot be ignored. Thus, there is a need for an alternate cryoprotectant. We reported previously that a novel cryopreservation reagent, StemCell Keep™ (SCK), was effective for cryopreserving human induced pluripotent stem cells (hiPSCs) by vitrification. Because hESCs and hiPSCs are not identical, the current study examined the use of SCK on hESCs...
May 9, 2017: Cell Transplantation
Eric Smalley
No abstract text is available yet for this article.
July 12, 2016: Nature Biotechnology
(no author information available yet)
We asked four stem cell scientists who recently started their labs or expanded their research programs to share their insights and approaches to obtaining funding. We present highlights from their interview responses here.
July 7, 2016: Cell Stem Cell
Maren Bredemeier, Philippos Edimiris, Mitra Tewes, Pawel Mach, Bahriye Aktas, Doreen Schellbach, Jenny Wagner, Rainer Kimmig, Sabine Kasimir-Bauer
BACKGROUND: Circulating tumor cells (CTC) are discussed to be an ideal surrogate marker for individualized treatment in metastatic breast cancer (MBC) since metastatic tissue is often difficult to obtain for repeated analysis. We established a nine gene qPCR panel to characterize the heterogeneous CTC population in MBC patients including epithelial CTC, their receptors (EPCAM, ERBB2, ERBB3, EGFR) CTC in Epithelial-Mesenchymal-Transition [(EMT); PIK3CA, AKT2), stem cell-like CTC (ALDH1) as well as resistant CTC (ERCC1, AURKA] to identify individual therapeutic targets...
July 5, 2016: Oncotarget
Marie Klintman, Richard Buus, Maggie Chon U Cheang, Amna Sheri, Ian E Smith, Mitch Dowsett
PURPOSE: The primary aim was to derive evidence for or against the clinical importance of several biologic processes in patients treated with neoadjuvant chemotherapy (NAC) by assessing expression of selected genes with prior implications in prognosis or treatment resistance. The secondary aim was to determine the prognostic impact in residual disease of the genes' expression. EXPERIMENTAL DESIGN: Expression levels of 24 genes were quantified by NanoString nCounter on formalin-fixed paraffin-embedded residual tumors from 126 patients treated with NAC and 56 paired presurgical biopsies...
May 15, 2016: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
Eva Schmelzer, Jörg C Gerlach
The creation of larger-scale three-dimensional tissue constructs depends on proper medium mass and gas exchange, as well as removal of metabolites, which cannot be achieved in conventional static two-dimensional petri dish culture. In cultures of tissue-density this problem can be addressed by decentral perfusion through artificial micro-capillaries. While the static medium exchange in petri dishes leads to metabolite peaks, perfusion culture provides a dynamic medium supply, thereby preventing non-physiological peaks...
2016: Methods in Molecular Biology
Motahere-Sadat Hashemi, Abbas Kiani Esfahani, Maryam Peymani, Alireza Shoaraye Nejati, Kamran Ghaedi, Mohammad Hossein Nasr-Esfahani, Hossein Baharvand
Zinc finger protein 521 is highly expressed in brain, neural stem cells and early progenitors of the human hematopoietic cells. Zfp521 triggers the cascade of neurogenesis in mouse embryonic stem cells through inducing expression of the early neuroectodermal genes Sox1, Sox3 and Pax6. Fndc5, a precursor of Irisin has inducing effects on the expression level of brain derived neurotrophic factor in hippocampus. Therefore, it is most likely that Fndc5 may play an important role in neural differentiation. To exhibit whether the expression of this protein is under regulation with Zfp521, we overexpressed Zfp521 in a stable transformants of mESCs expressing EGFP under control of Fndc5 promoter...
March 2016: Journal of Biosciences
(no author information available yet)
In an interview, we recently asked four leaders in the field to share their insights with us on the clinical outlook for hiPSCs. We present highlights from their responses here.
February 4, 2016: Cell Stem Cell
Pavlina Janovska, Lucie Poppova, Karla Plevova, Hana Plesingerova, Martin Behal, Marketa Kaucka, Petra Ovesna, Michaela Hlozkova, Marek Borsky, Olga Stehlikova, Yvona Brychtova, Michael Doubek, Michaela Machalova, Sivasubramanian Baskar, Alois Kozubik, Sarka Pospisilova, Sarka Pavlova, Vitezslav Bryja
PURPOSE: ROR1, a receptor in the noncanonical Wnt/planar cell polarity (PCP) pathway, is upregulated in malignant B cells of chronic lymphocytic leukemia (CLL) patients. It has been shown that the Wnt/PCP pathway drives pathogenesis of CLL, but which factors activate the ROR1 and PCP pathway in CLL cells remains unclear. EXPERIMENTAL DESIGN: B lymphocytes from the peripheral blood of CLL patients were negatively separated using RosetteSep (StemCell) and gradient density centrifugation...
January 15, 2016: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
Han-Wen Guo, Jia-Sin Yu, Shu-Han Hsu, Yau-Huei Wei, Oscar K Lee, Chen-Yuan Dong, Hsing-Wen Wang
Reduced nicotinamide adenine dinucleotide (NADH) fluorescence lifetime has been broadly used as a metabolic indicator for stem cell imaging. However, the direct relationship between NADH fluorescence lifetime and metabolic pathway and activity remains to be clarified. In this study, we measured the NADH fluorescence lifetime of human mesenchymal stem cells (hMSCs) as well as the metabolic indictors, such as adenosine triphosphate (ATP) level, oxygen consumption, and lactate release, up to 4 weeks under normal osteogenic differentiation and oxidative phosphorylation-attenuated/inhibited differentiation by oligomycin A (OA) treatment...
January 2015: Journal of Biomedical Optics
Hai-Sen Huang, Shuang Liu, Mei Yu, Xiao-Xin Luo, Wei-Hua Guo, Wei-Dong Tian
UNLABELLED: OBEJECTIVE: To construct miRNA-expressing plasmid vector for interfering the expression of beta-catenin in adipose-derived stem cells from Sprague-Dawley rats. METHODS: The double strands oligonucleotides of miR-expressing cassette were ligated into plasmid pSWH to generate pSWH-miR. Then the PCR product of enhanced green fluorescence protein (EGFP) open reading frame (ORF) fragment was inserted into BamH I and Xbo I digested pSWH-miR to generate pSWH-EGFP-miR...
September 2014: Sichuan da Xue Xue Bao. Yi Xue Ban, Journal of Sichuan University. Medical Science Edition
Manuel Sánchez-Castillo, David Ruau, Adam C Wilkinson, Felicia S L Ng, Rebecca Hannah, Evangelia Diamanti, Patrick Lombard, Nicola K Wilson, Berthold Gottgens
CODEX ( is a user-friendly database for the direct access and interrogation of publicly available next-generation sequencing (NGS) data, specifically aimed at experimental biologists. In an era of multi-centre genomic dataset generation, CODEX provides a single database where these samples are collected, uniformly processed and vetted. The main drive of CODEX is to provide the wider scientific community with instant access to high-quality NGS data, which, irrespective of the publishing laboratory, is directly comparable...
January 2015: Nucleic Acids Research
Nicola Maffulli, Umile Giuseppe Longo, Mattia Loppini, Alessandra Berton, Filippo Spiezia, Vincenzo Denaro
The purpose of this systematic review was to address the treatment of rotator cuff tears by applying tissue engineering approaches to improve tendon healing, specifically platelet rich plasma (PRP) augmentation, stem cells, and scaffolds. Our systematic search was performed using the combination of the following terms: "rotator cuff", "shoulder", "PRP", "platelet rich plasma", "stemcells", "scaffold", "growth factors", and "tissue engineering". No level I or II studies were found on the use of scaffolds and stem cells for rotator cuff repair...
2012: Stem Cells International
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