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  • [Colorectal Dis.] Isolation of internal and external sphincter progenitor cells from the human anal sphincter with or without radiotherapy.

    서울의대 / 손일태, 강성범*

  • 출처
    Colorectal Dis.
  • 등재일
    2018 Jul 26. doi: 10.1111/codi.14351. [Epub ahead
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    Abstract
    AIM:
    We aimed to isolate and propagate internal and external anal sphincter progenitor cells from the human anal sphincter, with or without radiotherapy, for tailored cell therapy of faecal incontinence.

    METHODS:
    Sphincter progenitor cells were isolated from normal internal and external anal sphincters collected from 10 patients with rectal cancer who had undergone abdominoperineal resection with (n = 6) or without (n = 4) preoperative chemoradiotherapy. The isolated cells and differentiated muscle fibres were identified using immunofluorescence assay, western blotting, and reverse transcription (RT)-PCR. The proliferation of progenitor cells with and without radiotherapy was compared by quantitative 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

    RESULTS:
    The immunofluorescence assay before differentiation confirmed that the internal anal sphincter progenitor cells expressed CD34 and neural-glial antigen 2 (NG2), whereas the external anal sphincter progenitor cells expressed CD34 and PAX7. After differentiation, the internal anal sphincter progenitor cells expressed desmin, calponin, and α-smooth muscle actin, whereas the external anal sphincter progenitor cells expressed desmin, myogenic factor 4, and myosin heavy chain. The differential expression profiles of both cell types were confirmed by western blotting and RT-PCR. MTT assays showed that the viability of internal and external anal sphincter progenitor cells was significantly lower in the radiotherapy group than that in the non-radiotherapy group.

    CONCLUSIONS:
    This study describes the differential harvest internal and external sphincter muscle progenitor cells from human anal sphincters. We confirm that radiotherapy decreases the viability of internal and external anal sphincter progenitor cells. This article is protected by copyright. All rights reserved.

     


    Author information

    Son IT1, Lee HS2, Ihn MH3, Lee KH4, Kim DW3, Lee KW5, Kim JS6, Kang SB3.
    1
    Department of Surgery, Seoul National University Bundang Hospital, Seongnam, South Korea.
    2
    Department of Surgery, Uijeongbu St. Mary's Hospital, Catholic University, Uijeongbu-si, South Korea.
    3
    Department of Pathology, Seoul National University College of Medicine, Seoul National University Bundang Hospital, Seongnam, South Korea.
    4
    Department of Radiology, Seoul National University College of Medicine, Seoul National University Bundang Hospital, Seongnam, South Korea.
    5
    Department of Hemato-Oncology, Seoul National University College of Medicine, Seoul National University Bundang Hospital, Seongnam, South Korea.
    6
    Department of Radiation Oncology, Seoul National University College of Medicine, Seoul National University Bundang Hospital, Seongnam, South Korea.

  • 키워드
    anal sphincter progenitor cell; cell viability; radiotherapy
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