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  • Ovid Technologies (Wolters Kluwer Health)  (3)
  • Takeda, Yusuke  (3)
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  • Ovid Technologies (Wolters Kluwer Health)  (3)
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  • 1
    Online Resource
    Online Resource
    Abstract 11725: Exosomes Secreted from TSG6-Rich Adipose-Derived Stem Cells Induce Neovascularization in Ischemic Limb
    Ovid Technologies (Wolters Kluwer Health) ; 2022
    In:  Circulation Vol. 146, No. Suppl_1 ( 2022-11-08)
    Details
    In: Circulation, Ovid Technologies (Wolters Kluwer Health), Vol. 146, No. Suppl_1 ( 2022-11-08)
    Abstract: Introduction: Therapeutic angiogenesis is attractive for tackling cardiovascular disease. Exosomes, which contain growth factors and micro RNAs, might be useful to develop cell-free therapeutic angiogenesis. Previously we have reported that CD271+ADRCs have excellent angiogenic activity. In this study, we investigated the in vivo angiogenic effects and mechanisms of CD271+ ADSC-derived exosomes (CD271+ Exo) using a mouse model of limb ischemia. Methods: We analyzed single-cell RNA-sequencing data of human stromal vascular fraction (SVF) cells. After normalization and clustering, CD271+ and CD271- cells were analyzed in DESeq2. Next, we established CD271+/- ADSCs from human SVF using FACS. After 48 hours of serum deprivation, conditioned media was collected. Exosomes were purified by magnetic isolation using phosphatidylserine/Tim4 interaction and confirmed by ELISA of exosomal markers. We labeled exosomes by PKH26 membrane dye and injected them into a mice model of limb ischemia. After 2 weeks, lectin perfused limb was harvested and analyzed by immunohistochemistry (Fig.A). Results: In single-cell transcriptome of human SVF, TSG6 was predominantly expressed in Lineage-CD34+ stromal cells. Furthermore, TSG6 was significantly upregulated in CD271+ SVF cells compared to CD271- SVF cells and other cluster cells (Fig.B). Consistently, TSG6 mRNA expression was upregulated in CD271+ ADSCs (Fig.C). We confirmed exosomal marker expression (CD63, CD9, and CD81) of isolated Exo (Fig.D). In exosome therapy, CD271+ Exo promotes neovascularization compared to CD271- Exo or PBS. Notably, there were more cells containing PKH labeling in the CD271+ Exo-treated group (Fig.E). Conclusions: This study reveals a novel mechanism by which CD271+ Exo promotes angiogenesis via TSG6. CD271+ Exo would be useful for cell-free therapeutic angiogenesis.
    Type of Medium: Online Resource
    ISSN: 0009-7322 , 1524-4539
    URL: Article
    DOI: 10.1161/circ.146.suppl_1.11725
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2022
    detail.hit.zdb_id: 1466401-X
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  • 2
    Online Resource
    Online Resource
    Abstract 10620: CD271 Expression Enhances Angiogenic Capacity in Human Adipose-Derived Stem Cells
    Ovid Technologies (Wolters Kluwer Health) ; 2022
    In:  Circulation Vol. 146, No. Suppl_1 ( 2022-11-08)
    Details
    In: Circulation, Ovid Technologies (Wolters Kluwer Health), Vol. 146, No. Suppl_1 ( 2022-11-08)
    Abstract: Introduction: Therapeutic angiogenesis using stem cell transplantation can be a novel strategy for ischemic disease. Recently, stem cells with gene modifications have been considered as an option to boost therapeutic efficacy. Surface receptor CD271 has emerged as a useful marker for proliferative or angiogenic cell isolation from human organs. We hypothesized CD271 expression on adipose-derived stem cells (ADSCs) would increase their angiogenic capacity. Purpose: We aimed to assess in vitro and in vivo angiogenic capacity of CD271-over expressed human ADSCs. Methods: ADSCs were established from human adipose tissue from healthy subjects and transfected with lentiviral vector for CD271 overexpression (OE-ADSCs). Vector transfection was performed with previously reported method using protamine. Vector without CD271 gene insert was used to prepare control (Cont-ADSCs, Fig.A ). Overexpression of CD271 was evaluated by qPCR and flow cytometry after cell selection using puromycin. For in vivo angiogenic capacity, ADSCs were prelabeled with PKH26 living cell dye, injected via intra-muscular injection into mice subjected to limb ischemia model, and evaluated by immunohistochemistry of ischemic limb (gastrocnemius). Results: We found that mRNA expression of CD271 was 10,000-fold higher in OE-ADSCs than in Cont-ADSCs. Consistently, over 90% of OE-ADSCs expressed CD271 protein. ( Fig.B ). Overexpression of CD271 on ADSCs increased mRNA expression of angiogenic factors such as VEGFA and CXCL12 . Furthermore, mRNA expression of TIMP2, anti-angiogenic factor, was significantly decreased in OE-ADSCs ( Fig.C ). In cell therapy, we demonstrated that administration of OE-ADSCs enhanced in vivo angiogenic capacity evaluated by capillary density. Notably, ADSC engraftment capacity was also augmented in OE-ADSCs ( Fig.D ). Conclusion: These findings highlight the important role of CD271 expression on ADSCs for their angiogenic gene profile and cell therapy effects.
    Type of Medium: Online Resource
    ISSN: 0009-7322 , 1524-4539
    URL: Article
    DOI: 10.1161/circ.146.suppl_1.10620
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2022
    detail.hit.zdb_id: 1466401-X
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  • 3
    Online Resource
    Online Resource
    Abstract 402: Human CD271-positive Adipose Derived Stem Cells are the Angiogenic Subset Decreased by Donor Insulin Resistance
    Ovid Technologies (Wolters Kluwer Health) ; 2020
    In:  Circulation Research Vol. 127, No. Suppl_1 ( 2020-07-31)
    Details
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 127, No. Suppl_1 ( 2020-07-31)
    Abstract: Introduction: Type 2 diabetes mellitus (T2DM) is an important risk factor for cardiovascular diseases (CVDs). Recently, cell therapy using adipose-derived stem cells (ADSCs) has emerged as an attractive therapy for severe CVDs because of their angiogenic potentials. However, whether and how T2DM would impair human ADSC angiogenic capacity is still uncertain. We previously reported that CD31 - CD34 + CD271 + ADSCs (CD271 + ADSCs) were specifically decreased in adipose tissue of T2DM patients. Therefore, we aimed to investigate the angiogenic capacity of CD271 + ADSCs. Furthermore, we evaluate which patients’ parameters regard as T2DM would decrease the amount of CD271 + ADSCs. Methods and Results: Human CD45 - CD34 + CD31 - ADSCs were obtained from subcutaneous adipose tissue of healthy donors, separated into CD271 + and CD271 - subsets by FACS, and cultured. Both subsets of ADSCs were assessed gene expression profile by microarray. Microarray analysis and validation PCR elucidated that PI3K/Akt/mTOR pathway was significantly up-regulated in CD271 + ADSCs compared to in CD271 - ADSCs. ( p 〈 0.05). Then, we compared in vivo angiogenic capacity in xenograft experiments of nude mice subjected to hindlimb ischemia. Angiogenesis was evaluated histologically using perfused lectin (capillary density) at day 14. Cell therapy using CD271 + ADSCs demonstrated about 3-fold more lectin + capillaries compared to CD271 - ADSCs or PBS injection ( p 〈 0.005, n = 5 / group). Next, we established cultured ADSCs obtained from CD271 knock-out mice (KO-ADSCs) and compared their angiogenic capacity with those from WT mice. Consistently, KO-ADSCs demonstrated impaired in vivo angiogenic capacity ( p 〈 0.005, n = 5 / group). Finally, we collected 23 samples of adipose tissue obtained from CVD patients and evaluated the frequency of CD271 + ADSCs in CD45 - CD34 + CD31 - ADSCs. Among studied parameters, HOMA-IR, an index of insulin resistance, was negatively correlated with the frequency of CD271+ ADSCs ( r = -0.64, p 〈 0.005). Conclusions: Human CD271 + ADSCs demonstrated enhanced in vivo angiogenic capacity with higher mTOR expression. Donor insulin resistance might decrease this regenerative subset of ADSCs. These findings would be critical for development and improvement of ADSC therapy.
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
    URL: Article
    DOI: 10.1161/res.127.suppl_1.402
    RVK:
    XA 10000
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2020
    detail.hit.zdb_id: 1467838-X
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