OBJECTIVE: To purify human muscle-derived cells (hMDCs) that could be used for managing urinary incontinence, erectile dysfunction and for bladder reconstitution. MATERIALS AND METHODS: hMDCs isolated by a modified preplate technique (MPT) from skeletal muscles of limbs were purified by CD34 antibody and magnetic Dynabeads cell selection system (MDCSS). The growth-doubling time of the hMDCs and purified hMDCs was measured. The purified hMDCs were characterized by flow cytometry, immunofluorescence and confocal laser scanning microscopy. RESULTS: The growth-doubling time of hMDCs was approximately 24 h, which increased to 35 h after purifying using the MDCSS. There were scanty fibroblasts after purification and the MDCSS-purified hMDCs were identified by high expression of stem cell markers and myoblast markers. The expression proportion of the stem cell marker CD34 and myoblast marker CD56 in the hMDCs was higher after purification (MDCSS) than before (MPT), at 32.13% vs 4.12% and 21.56% vs 8.60%, respectively. CONCLUSIONS: The purification of hMDCs showing high expression of stem cell and myoblast markers is feasible. These purified hMDCs could potentially be used for urological regeneration.
OBJECTIVE: To purify human muscle-derived cells (hMDCs) that could be used for managing urinary incontinence, erectile dysfunction and for bladder reconstitution. MATERIALS AND METHODS: hMDCs isolated by a modified preplate technique (MPT) from skeletal muscles of limbs were purified by CD34 antibody and magnetic Dynabeads cell selection system (MDCSS). The growth-doubling time of the hMDCs and purified hMDCs was measured. The purified hMDCs were characterized by flow cytometry, immunofluorescence and confocal laser scanning microscopy. RESULTS: The growth-doubling time of hMDCs was approximately 24 h, which increased to 35 h after purifying using the MDCSS. There were scanty fibroblasts after purification and the MDCSS-purified hMDCs were identified by high expression of stem cell markers and myoblast markers. The expression proportion of the stem cell marker CD34 and myoblast marker CD56 in the hMDCs was higher after purification (MDCSS) than before (MPT), at 32.13% vs 4.12% and 21.56% vs 8.60%, respectively. CONCLUSIONS: The purification of hMDCs showing high expression of stem cell and myoblast markers is feasible. These purified hMDCs could potentially be used for urological regeneration.
Authors: Natalia Rozwadowska; Maria Sikorska; Katarzyna Bozyk; Klaudia Jarosinska; Anna Cieciuch; Sonia Brodowska; Marcin Andrzejczak; Maria Siemionow Journal: Am J Stem Cells Date: 2022-02-15