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Isolation of internal and external sphincter progenitor cells from the human anal sphincter with or without radiotherapy
Author(s) -
Son I. T.,
Lee H. S.,
Ihn M. H.,
Lee K. H.,
Kim D.W.,
Lee K.W.,
Kim J.S.,
Kang S.B.
Publication year - 2019
Publication title -
colorectal disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.029
H-Index - 89
eISSN - 1463-1318
pISSN - 1462-8910
DOI - 10.1111/codi.14351
Subject(s) - progenitor cell , medicine , urethral sphincter , external anal sphincter , cd34 , progenitor , internal anal sphincter , radiation therapy , pathology , stem cell , urology , anal canal , biology , rectum , microbiology and biotechnology , urethra
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 polymerase chain reaction ( 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 CD 34 and neural‐glial antigen 2 ( NG 2), whereas the external anal sphincter progenitor cells expressed CD 34 and PAX 7. 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 nonradiotherapy 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.