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PTN Nghiên cứu và Ứng dụng Tế bào gốc

Category: Bài báo ngoài nước

  • Isolation and culture of neural stem cells from murine foetal brain

    Isolation and culture of neural stem cells from murine foetal brain

    Nhung Hai Truong, Nhung Thi-Hong Dinh, Dung Minh Le, Linh Thuy Nguyen, Thanh Thai Lam, Ngoc Kim Phan and Phuc Van Pham*

    Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh city, Vietnam

    Abstract

    There are many mysteries of the nervous system and neurodegenerative diseases. Evidences showed that neural stem cells (NSCs) play a crucial role in recovering nervous system, although this was not sufficient due to lack of cell number. This research was carried out to isolate and culture NSCs from murine foetal brain and our NSCs were cultured as floating and multicellular neurospheres. The neurosphere has been consider as an excellent tool to investigate the differentiation and proliferation of NSCs. Propagation of NSCs in vitro was essential for understanding neural cell’s fate and supplying a promising source for cell therapy for neurodegenerative disease. Serum-free medium with mitogens, N2 and B27 supplement was appropriate for NSC culture. To characterize candidate cells, we assessed stemness and stem cell markers by sphere formation assay, differentiation assay, and immunocytochemistry. The results showed that NSCs could develop as floating spheres; they expressed stem cell- like characteristics and popular marker of neural stem cells-nestin, and differentiate into nerve cells. This population of cells was a promising source to studying neural stem cells and the treatment of neurodegenerative disease.

    Keywords: Neural stem cells; neurospheres; murine foetal brain; neurodegenerative disease; stem cells

    To cite this article: Truong NH, NTH Dinh, DM Le, LT Nguyen, TT Lam, NK Phan and PV Pham, 2014. Isolation and culture of neural stem cells from murine foetal brain. Res. Opin. Anim. Vet. Sci., 4(1), 24-29.

    http://roavs.com/Abstract/Issue-1-2014/24-29.htm

  • In vitro evaluation of the effects of human umbilical cord extracts on human fibroblasts, keratinocy

    In Vitro Cell Dev Biol Anim. 2013 Oct 26. [Epub ahead of print]

    In vitro evaluation of the effects of human umbilical cord extracts on human fibroblasts, keratinocytes, and melanocytes.

    Abstract

    Skin aging is the result of internal and external factors. So-called photoaging has been identified as the major factor in skin aging. Effects of photoaging include inhibition of fibroblast and keratinocyte proliferation as well as collagen and fibronectin expression, while activating expression of collagenases such as matrix metalloproteinase-1. Previous studies have shown that extracts or products from human placenta significantly improve skin aging and chronic wound healing. However, there are few studies of umbilical cord extracts. Therefore, this study aimed to evaluate the effects of umbilical cord extract-derived formulae on three kinds of skin cells including fibroblasts, keratinocytes, and melanocytes. We prepared 20 formulae from intracellular umbilical cord extracts, extracellular umbilical cord extracts, and umbilical cord-derived stem cell extracts, as well as five control formulae. We evaluated the effects of the 25 formulae on fibroblast and keratinocyte proliferation, and expression of collagen I, fibronectin, and matrix metalloproteinase-1 in fibroblasts and tyrosinase in melanocytes. The results showed that 7.5% formula 35 was the most effective formula for promotion of fibroblast and keratinocyte proliferation. At this concentration, formula 35 also induced collagen expression and inhibited matrix metalloproteinase-1 expression at the transcriptional level. However, this formula had no effect on tyrosinase expression in melanocytes. These results demonstrate that umbilical cord extracts can serve as an attractive source of proteins for skincare and chronic wound healing products.

    PMID:
    24163162
    [PubMed – as supplied by publisher]

  • Pegylated dendrimer and its effect in fluorouracil loading and release for enhancing antitumor activ

    J Biomed Nanotechnol. 2013 Feb;9(2):213-20.

    Pegylated dendrimer and its effect in fluorouracil loading and release for enhancing antitumor activity.

    Abstract

    Dendrimer, a new class of hyper-branched polymer with predetermined molecular weight, is being received much attention in nano biomedical applications such as anticancer drug delivery, gene therapy, disease diagnosis and etc. In this study, polyamidoamine (PAMAM)-based dendrimer generation 3.0 (G 3.0) was synthesized and subsequently pegylated. Obtained results showed that pegylation degree of the dendrimer was around 31% for its external amine groups. TEM image of the pegylated dendrimer exhibited spherical shape and nano sizes ranging from 30 to 40 nm. The fluorouracil (5-FU)-loaded pegylated dendrimer showed a slow release profile of the drug. In vitro study, at the primary screening concentration of 100 microg/mL, the PAMAM dendrimer presented higher toxicity in MCF-7 cells as compared to its pegylated counterpart. Meanwhile, the (5-FU)-loaded pegylated dendrimer exhibited the antiproliferative activity against the cell line with the IC50 of 9.92 +/- 0.19 microg/mL. In vivo tumor xenograft study, we succeeded in generating MCF-7 cells-derived cancer tumors on mice that was well-confirmed by using flow cytometer assay. The 5-FU encapsulated pegylated dendrimer exhibited a significant decrement in volume of the tumors which was generated by MCF-7 cancer cells.

    PMID:
    23627047
    [PubMed – indexed for MEDLINE]

  • Pegylated dendrimer and its effect in fluorouracil loading and release for enhancing antitumor activ

    J Biomed Nanotechnol. 2013 Feb;9(2):213-20.

    Pegylated dendrimer and its effect in fluorouracil loading and release for enhancing antitumor activity.

    Abstract

    Dendrimer, a new class of hyper-branched polymer with predetermined molecular weight, is being received much attention in nano biomedical applications such as anticancer drug delivery, gene therapy, disease diagnosis and etc. In this study, polyamidoamine (PAMAM)-based dendrimer generation 3.0 (G 3.0) was synthesized and subsequently pegylated. Obtained results showed that pegylation degree of the dendrimer was around 31% for its external amine groups. TEM image of the pegylated dendrimer exhibited spherical shape and nano sizes ranging from 30 to 40 nm. The fluorouracil (5-FU)-loaded pegylated dendrimer showed a slow release profile of the drug. In vitro study, at the primary screening concentration of 100 microg/mL, the PAMAM dendrimer presented higher toxicity in MCF-7 cells as compared to its pegylated counterpart. Meanwhile, the (5-FU)-loaded pegylated dendrimer exhibited the antiproliferative activity against the cell line with the IC50 of 9.92 +/- 0.19 microg/mL. In vivo tumor xenograft study, we succeeded in generating MCF-7 cells-derived cancer tumors on mice that was well-confirmed by using flow cytometer assay. The 5-FU encapsulated pegylated dendrimer exhibited a significant decrement in volume of the tumors which was generated by MCF-7 cancer cells.

    PMID:
    23627047
    [PubMed – indexed for MEDLINE]

  • Activated platelet-rich plasma improves adipose-derived stem cell transplantation efficiency in…

    Activated platelet-rich plasma improves adipose-derived stem cell transplantation efficiency in injured articular cartilage

    Phuc Van Pham, Khanh Hong-Thien Bui, Dat Quoc Ngo, Ngoc Bich Vu, Nhung Hai Truong, Nhan Lu-Chinh Phan, Dung Minh Le, Triet Dinh Duong, Thanh Duc Nguyen, Vien Tuong Le and Ngoc Kim Phan

    •  

    Email: Phuc Van Pham pvphuc@hcmuns.edu.vn – Khanh Hong-Thien Bui khanhbui1969@yahoo.com – Dat Quoc Ngo quocdat_yds@yahoo.com – Ngoc Bich Vu vbngoc@hcmus.edu.vn – Nhung Hai Truong thnhung@hcmuns.edu.vn – Nhan Lu-Chinh Phan plcnhan@hcmus.edu.vn – Dung Minh Le leminhdung90@gmail.com – Triet Dinh Duong bsduongdinhtriet@yahoo.com.vn – Thanh Duc Nguyen dr.ducthanh@yahoo.com – Vien Tuong Le tuongvien2007@yahoo.com.vn – Ngoc Kim Phan pkngoc@hcmuns.edu.vn

    Stem Cell Research & Therapy 2013, 4:91 doi:10.1186/scrt277

    Published: 1 August 2013

    Abstract (provisional)

    Introduction

    Adipose-derived stem cells (ADSCs) have been isolated, expanded, and applied in the treatment of many diseases. ADSCs have also been used to treat injured articular cartilage. However, there is controversy regarding the treatment efficiency. We considered that ADSC transplantation with activated platelet-rich plasma (PRP) may improve injured articular cartilage compared with that of ADSC transplantation alone. In this study, we determined the role of PRP in ADSC transplantation to improve the treatment efficiency.

    Methods

    ADSCs were isolated and expanded from human adipose tissue. PRP was collected and activated from human peripheral blood. The effects of PRP were evaluated in vitro and in ADSC transplantation in vivo. In vitro, the effects of PRP on ADSC proliferation, differentiation into chondrogenic cells, and inhibition of angiogenic factors were investigated at three concentrations of PRP (10%, 15%, and 20%). In vivo, ADSCs pre-treated with or without PRP were transplanted into murine models of injured articular cartilage.

    Results

    PRP promoted ADSC proliferation and differentiation into chondrogenic cells that strongly expressed collagen II, Sox9, and aggrecan. Moreover, PRP inhibited expression of the angiogenic factor vascular endothelial growth factor. As a result, PRP pre-treated ADSCs improved healing of injured articular cartilage in murine models compared with that of untreated ADSCs.

    Conclusions

    Pre-treatment of ADSCs with PRP is a simple method to efficiently apply ADSCs in cartilage regeneration. This study provides an important step toward the use of autologous ADSCs in the treatment of injured articular cartilage.

    scrt

  • Stimulation of Allogenic lymphocytes by Dendritic Cells Derived from Human Umbilical Cord Blood …

    Stimulation of Allogenic lymphocytes by Dendritic Cells Derived from Human Umbilical Cord Blood Fused with Breast Cancer Stem Cells

    Sinh Truong Nguyen 1, Viet Quoc Pham1, Ngoc Kim Phan1 and Phuc Van Pham1*

    1Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Vietnam.

     

    Aims: Cancer stem cells (CSCs) are cancer cells that possess characteristics associated with normal stem cells. CSCs represent a minor subset of cells in the tumor and are thought to be the reason for the initiation of disease, resistance to cancer treatment, and the occurrence of metastasis. Therefore, breast cancer stem cells (BCSCs) targeting therapies are considered as the promising therapy for breast cancer treatment. This research aims to evaluate the tumor associated antigens presentation of dendritic cells fused with breast cancer stem cells in dendritic cells based therapies.
    Methodology: Human breast cancer stem cells are isolated from malignant breast tumors by enrich culture and fluorescent activated cell sorting. Human dendritic cells are isolated from umbilical cord blood by culturing CD14 monocytes in induced medium. Electrocution is used to fuse breast cancer stem cells and dendritic cells. Fusion cells are used to evaluate functions of dendritic cells (DCs) and also to stimulate T cells.
    Results: These findings indicate that fusion cells have the ability to present the antigens of breast cancer stem cell to T-cells, and regarding functionality.
    Conclusion: They appear to be very good candidates for antitumor vaccine in breast cancer.

     

    Keywords :

    Antitumor vaccine; breast cancer stem cell; cell fusion; dendritic cell; tumor-associated antigens; T-cell proliferation.

     

    http://www.sciencedomain.org/abstract.php?iid=174&id=9&aid=823

     

     

  • Transplantation of non-expanded adipose stromal vascular fraction and platelet rich plasma for…

    Transplantation of non-expanded adipose stromal vascular fraction and platelet rich plasma for articular cartilage injury treatment in mice model

    Phuc Pham Van, Khanh Bui-Hong-Thien, Dat Ngo-Quoc, Lam Khuat-Tan, and Ngoc Phan-Kim

    1Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Ho Chi Minh city, Vietnam

    2University of Medical Center, Ho Chi Minh University of Medicine and Pharmacy, Ho Chi Minh City, Vietnam

    3Department of Pathology, University ofMedicine and Pharmacy, Ho Chi Minh City, Vietnam Corresponding author: pvphuc@hcmuns.edu.vn

    Received 13 August 2012; Accepted 25 December 2012

    Abstract

    Stromal vascular fraction (SVF) combined with platelet rich plasma (PRP) commonly used in preclinical and clinical osteoarthritis as well as articular cartilage injury treatment. However, this therapy has not been carefully evaluated the safety and efficacy. This research aims to assess the safety and efficacy of SVF combined with PRP transplantation. Ten samples of SVFs and PRPs from donors were used in this research. About safety, we evaluate the expression of some genes related to tumor formation such as Oct-4, Nanog, SSEA3, SSEA4 by RT-PCR, flow cytometry and tumor formation when injected in NOD/SCID mice. About efficacy, SVF was injected with PRP into murine joint that caused joint failure. The results showed that SVFs are negative with Oct-4, Nanog, SSEA-3, SSEA-4 as well as can not cause tumors in mice. SVFs combined with PRP can improve the joint regeneration in mice. These results proved that SVFs combined with PRP transplantation is a promising therapy for articular cartilage injury treatment.

    Keywords: Adipose stromal vascular fraction, joint failure, osteoarthritis, platelet rich plasma, mesenchymal stem cells.

     

    Online at:
    http://www.hindawi.com/journals/jme/aip/832396/

  • Doxorubicin and 5-fluorouracil resistant hepatic cancer cells demonstrate stem-like properties

    Doxorubicin and 5-fluorouracil resistant hepatic cancer cells demonstrate stem-like properties. Cytotechnology, 2012. DOI: 10.1007/s10616-012-9511-9.

    Ngoc Bich Vu, Tam Thanh Nguyen, Long Cong-Duy Tran, Cong Dinh Do, Bac Hoang Nguyen, Ngoc Kim Phan and Phuc Van Pham

    Abstract

    The efficacy of hepatocellular carcinoma (HCC) treatment is very low because of the high percentage of recurrence and resistance to anticancer agents. Hepatic cancer stem cells (HCSCs) are considered the origin of such recurrence and resistance. Our aim was to evaluate the stemness of doxorubicin and 5-fluorouracil resistant hepatic cancer cells and establish the new method to isolate the HCSCs from primary cultured HCC tumors. HCC biopsies were used to establish primary cultures. Then, primary cells were selected for HCSCs by culture in medium supplemented with doxorubicin (0, 0.1, 0.25, 0.5 or 1 μg/mL), 5-fluorouracil (0, 0.1, 0.25, 0.5 or 1 μg/mL) or their combination. Selection was confirmed by detection of HCSC markers such as CD133, CD13, CD90, and the side population was identified by rhodamine 123 efflux. The cell population with the strongest expression of these markers was used to evaluate the cell cycle, gene expression profile, tumor sphere formation, marker protein expression, and in vivo tumorigenesis. Selective culture of primary cells in medium supplemented with 0.5 μg/mL doxorubicin and 1 μg/mL 5-fluorouracil selected cancer cells with the highest stemness properties. Selected cells strongly expressed CD13, CD133, CD90, and CD326, efflux rhodamine 123 and formed tumor spheres in suspension. Moreover, selected cells were induced to differentiate into cells with high expression of CD19 and AFP (alpha-fetoprotein), and importantly, could form tumors in NOD/SCID mice upon injection of 1 × 105 cells/mouse. Selective culture with doxorubicin and 5-fluorouracil will enrich HCSCs, is an easy method to obtain HCSCs that can be used to develop better therapeutic strategies for patients with HCC, and particularly HCSC-targeting therapy.

    springer pubmed

  • c-Maf plays a crucial role for the definitive erythropoiesis that accompanies erythroblastic island

    c-Maf plays a crucial role for the definitive erythropoiesis that accompanies erythroblastic island formation in the fetal liver

    1. Manabu Kusakabe1,2,
    2. Kazuteru Hasegawa1,
    3. Michito Hamada1,
    4. Megumi Nakamura1,
    5. Takayuki Ohsumi1,
    6. Hirona Suzuki1,
    7. Tran Thi Nhu Mai3,
    8. Takashi Kudo1,
    9. Kazuhiko Uchida4,
    10. Haruhiko Ninomiya2,
    11. Shigeru Chiba2, and
    12. Satoru Takahashi1

    – Author Affiliations


    1. 1Department of Anatomy and Embryology, Institute of Basic Medical Sciences, and

    2. 2Department of Hematology, Institute of Clinical Medicine, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan;

    3. 3Laboratory of Stem Cell Research and Application, University of Science, Ho Chi Minh City, Vietnam; and

    4. 4Department of Molecular Biological Oncology, Institute of Basic Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan

      5. pubmedBlood

  • Suppression of human breast tumors in NOD/SCID mice by CD44 shRNA gene therapy combined…

    Suppression of human breast tumors in NOD/SCID mice by CD44 shRNA gene therapy combined with doxorubicin treatment

    Pham PV, Vu NB, Duong TT, Nguyen TT, Truong NH, Phan NLC, Vuong TG, Pham VQ, Nguyen HM, Nguyen KT, Nguyen NT, Nguyen KG, Khat LT, Le DV, Truong KD, Phan NK

    Published Date May 2012 Volume 2012:5 Pages 77 – 84

    DOI: http://dx.doi.org/10.2147/OTT.S30609

     

    Phuc Van Pham1, Ngoc Bich Vu1, Thuy Thanh Duong1, Tam Thanh Nguyen1, Nhung Hai Truong1, Nhan Lu Chinh Phan1, Tue Gia Vuong1, Viet Quoc Pham1, Hoang Minh Nguyen1, Kha The Nguyen1, Nhung Thi Nguyen1, Khue Gia Nguyen1, Lam Tan Khat1, Dong Van Le2, Kiet Dinh Truong1, Ngoc Kim Phan1

    1Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, HCM City, 2Military Medical University, Ha Noi, Vietnam

    Background: Breast cancer stem cells with a CD44+CD24– phenotype are the origin of breast tumors. Strong CD44 expression in this population indicates its important role in maintaining the stem cell phenotype. Previous studies show that CD44 down-regulation causes CD44+CD24– breast cancer stem cells to differentiate into non-stem cells that are sensitive to antitumor drugs and lose many characteristics of the original cells. In this study, we determined tumor suppression in non-obese severe combined immunodeficiency mice using CD44 shRNA therapy combined with doxorubicin treatment.
    Methods: Tumor-bearing non-obese severe combined immunodeficiency mice were established by injection of CD44+CD24– cells. To track CD44+CD24– cells, green fluorescence protein was stably transduced using a lentiviral vector prior to injection into mice. The amount of CD44 shRNA lentiviral vector used for transduction was based on CD44 down-regulation by in vitro CD44 shRNA transduction. Mice were treated with direct injection of CD44 shRNA lentiviral vector into tumors followed by doxorubicin administration after 48 hours. The effect was evaluated by changes in the size and weight of tumors compared with that of the control.
    Results: The combination of CD44 down-regulation and doxorubicin strongly suppressed tumor growth with significant differences in tumor sizes and weights compared with that of CD44 down-regulation or doxorubicin treatment alone. In the combination of CD44 down-regulation and doxorubicin group, the tumor weight was significantly decreased by 4.38-fold compared with that of the control group.
    Conclusion: These results support a new strategy for breast cancer treatment by combining gene therapy with chemotherapy.

    Keywords: breast cancer, breast cancer stem cells, CD44, doxorubicin, gene therapy

    dove

    pubmed