Claudia Gil-Sanchis1, Irene Cervelló1, Satish Khurana2, Amparo Faus1, Catherine Verfaillie2, Carlos Simón3. 1. Fundación IVI-Instituto Universitario IVI-Universidad de Valencia, INCLIVA, Valencia, Spain. 2. Stem Cell Institute KU Leuven, Leuven, Belgium. 3. Fundación IVI-Instituto Universitario IVI-Universidad de Valencia, INCLIVA, Valencia, Spain; Department of Obstetrics and Gynecology, School of Medicine, Valencia University, Valencia, Spain; Department of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford University, Stanford, California. Electronic address: Carlos.Simon@ivi.es.
Abstract
OBJECTIVE: To study the involvement of seven types of bone marrow-derived cells (BMDCs) in the endometrial regeneration in mice after total body irradiation. DESIGN: Prospective experimental animal study. SETTING: University research laboratories. ANIMAL(S): β-Actin-green fluorescent protein (GFP) transgenic C57BL/6-Tg (CAG-EGFP) and C57BL/6J female mice. INTERVENTION(S): The BMDCs were isolated from CAG-EGFP mice: unfractionated bone marrow cells, hematopoietic progenitor cells, endothelial progenitor cells (EPCs), and mesenchymal stem cells (MSCs). In addition three murine GFP(+) cell lines were used: mouse Oct4 negative BMDC multipotent adult progenitor cells (mOct4(-)BM-MAPCs), BMDC hypoblast-like stem cells (mOct4(+) BM-HypoSCs), and MSCs. All cell types were injected through the tail vein of 9 Gy-irradiated C57BL/6J female mice. MAIN OUTCOME MEASURE(S): Flow cytometry, cell culture, bone marrow transplantation assays, histologic evaluation, immunohistochemistry, proliferation, apoptosis, and statistical analysis. RESULT(S): After 12 weeks, histologic analysis revealed that uteri of mice with mOct4(-)BM-MAPCs and MSC line were significantly smaller than uteri of mice with uncultured BMDCs or mOct4(+) BM-HypoSCs. The percentage of engrafted GFP(+) cells ranged from 0.13%-4.78%. Expression of Ki-67 was lower in all uteri from BMDCs treated mice than in the control, whereas TUNEL(+) cells were increased in the EPCs and mOct4(+)BM-HypoSCs groups. CONCLUSION(S): Low number of some BMDCs can be found in regenerating endometrium, including stromal, endotelial, and epithelial compartments. Freshly isolated MSCs and EPCs together with mOct4(+) BM-HypoSCs induced the greatest degree of regeneration, whereas culture isolated MSCs and mOct4(-)BM-MAPCs transplantation may have an inhibitory effect on endometrial regeneration.
OBJECTIVE: To study the involvement of seven types of bone marrow-derived cells (BMDCs) in the endometrial regeneration in mice after total body irradiation. DESIGN: Prospective experimental animal study. SETTING: University research laboratories. ANIMAL(S): β-Actin-green fluorescent protein (GFP) transgenic C57BL/6-Tg (CAG-EGFP) and C57BL/6J female mice. INTERVENTION(S): The BMDCs were isolated from CAG-EGFP mice: unfractionated bone marrow cells, hematopoietic progenitor cells, endothelial progenitor cells (EPCs), and mesenchymal stem cells (MSCs). In addition three murine GFP(+) cell lines were used: mouseOct4 negative BMDC multipotent adult progenitor cells (mOct4(-)BM-MAPCs), BMDC hypoblast-like stem cells (mOct4(+) BM-HypoSCs), and MSCs. All cell types were injected through the tail vein of 9 Gy-irradiated C57BL/6J female mice. MAIN OUTCOME MEASURE(S): Flow cytometry, cell culture, bone marrow transplantation assays, histologic evaluation, immunohistochemistry, proliferation, apoptosis, and statistical analysis. RESULT(S): After 12 weeks, histologic analysis revealed that uteri of mice with mOct4(-)BM-MAPCs and MSC line were significantly smaller than uteri of mice with uncultured BMDCs or mOct4(+) BM-HypoSCs. The percentage of engrafted GFP(+) cells ranged from 0.13%-4.78%. Expression of Ki-67 was lower in all uteri from BMDCs treated mice than in the control, whereas TUNEL(+) cells were increased in the EPCs and mOct4(+)BM-HypoSCs groups. CONCLUSION(S): Low number of some BMDCs can be found in regenerating endometrium, including stromal, endotelial, and epithelial compartments. Freshly isolated MSCs and EPCs together with mOct4(+) BM-HypoSCs induced the greatest degree of regeneration, whereas culture isolated MSCs and mOct4(-)BM-MAPCs transplantation may have an inhibitory effect on endometrial regeneration.
Authors: Hilary O D Critchley; Elnur Babayev; Serdar E Bulun; Sandy Clark; Iolanda Garcia-Grau; Peter K Gregersen; Aoife Kilcoyne; Ji-Yong Julie Kim; Missy Lavender; Erica E Marsh; Kristen A Matteson; Jacqueline A Maybin; Christine N Metz; Inmaculada Moreno; Kami Silk; Marni Sommer; Carlos Simon; Ridhi Tariyal; Hugh S Taylor; Günter P Wagner; Linda G Griffith Journal: Am J Obstet Gynecol Date: 2020-07-21 Impact factor: 10.693