P-1028: Human umbilical cord blood stem cells may incorporate into uterus of NOD/SCID mice by intravenous injection

Abstract

ObjectiveThe uterine endometrium is renewed in every menstrual cycle and it should be thought that endometrium may contain a lot of endometrial stem cells. These endogeneous stem cells can rapidly regenerate the endometrium to support pregnancy by hormone regulation. However, disorders of the endometrium may avoid the generation and growth of endometrial cells and finally lead infertility. From recent publication, donor-derived endometrial cells were detected in endometrial biopsy samples from all bone marrow recipients (Taylor, 2004). So, the aim of this study was to evaluate the possibility that human umbilical cord blood (UCB) stem cells could be differentiated into endometrial stem cells after transplantation.DesignTransplantation of human UCB stem cells into NOD/SCID mice by intravenous injection into a lateral tail vein.Materials and methodsUCB was obtained from Public Umbilical Cord Blood Bank (I-cord, Chabiotech Co., Inc., Seoul, Korea) under approval of IRB of CHA General Hospital, Seoul, Korea. Mononuclear cells (MNCs) were isolated following density gradient separation with Ficoll and CD34+ cells were positively selected directly from MNCs by immunomagnetic cell separation. Female NOD/SCID mice, 8 to 12 weeks of age, were maintained under specific pathogen-free conditions. MNCs (2x107) or CD34+ cells (2x105) derived from UCB were transplanted into NOD/SCID mice by intravenous injection into a lateral tail vein. Two or 4 weeks after transplantation, uterus of mice was recovered and sampled for PCR of human specific primer set (the α-satellite DNA on human chromosome 17) and immunohistochemistry using anti-human nuclear antigen antibody (Chemicon) and peroxidase/DAB staining kit (DAKO).ResultsAt 2 weeks, signal of human specific primer set was not detected in uterus of MNCs and CD34+ cells-transferred mice. However, strong signals were detected in uterus of all of transplanted mice after 4 weeks. Immunohistochemistry revealed human cells originated from MNCs and CD34+ cells in stroma and glandular epithelium of mouse uterus.ConclusionThese preliminary finding suggest that human UCB stem cells can incorporate into the uterus by intravenous injection and may sustain. Functional regeneration to endometrial cells will be further investigated. ObjectiveThe uterine endometrium is renewed in every menstrual cycle and it should be thought that endometrium may contain a lot of endometrial stem cells. These endogeneous stem cells can rapidly regenerate the endometrium to support pregnancy by hormone regulation. However, disorders of the endometrium may avoid the generation and growth of endometrial cells and finally lead infertility. From recent publication, donor-derived endometrial cells were detected in endometrial biopsy samples from all bone marrow recipients (Taylor, 2004). So, the aim of this study was to evaluate the possibility that human umbilical cord blood (UCB) stem cells could be differentiated into endometrial stem cells after transplantation. The uterine endometrium is renewed in every menstrual cycle and it should be thought that endometrium may contain a lot of endometrial stem cells. These endogeneous stem cells can rapidly regenerate the endometrium to support pregnancy by hormone regulation. However, disorders of the endometrium may avoid the generation and growth of endometrial cells and finally lead infertility. From recent publication, donor-derived endometrial cells were detected in endometrial biopsy samples from all bone marrow recipients (Taylor, 2004). So, the aim of this study was to evaluate the possibility that human umbilical cord blood (UCB) stem cells could be differentiated into endometrial stem cells after transplantation. DesignTransplantation of human UCB stem cells into NOD/SCID mice by intravenous injection into a lateral tail vein. Transplantation of human UCB stem cells into NOD/SCID mice by intravenous injection into a lateral tail vein. Materials and methodsUCB was obtained from Public Umbilical Cord Blood Bank (I-cord, Chabiotech Co., Inc., Seoul, Korea) under approval of IRB of CHA General Hospital, Seoul, Korea. Mononuclear cells (MNCs) were isolated following density gradient separation with Ficoll and CD34+ cells were positively selected directly from MNCs by immunomagnetic cell separation. Female NOD/SCID mice, 8 to 12 weeks of age, were maintained under specific pathogen-free conditions. MNCs (2x107) or CD34+ cells (2x105) derived from UCB were transplanted into NOD/SCID mice by intravenous injection into a lateral tail vein. Two or 4 weeks after transplantation, uterus of mice was recovered and sampled for PCR of human specific primer set (the α-satellite DNA on human chromosome 17) and immunohistochemistry using anti-human nuclear antigen antibody (Chemicon) and peroxidase/DAB staining kit (DAKO). UCB was obtained from Public Umbilical Cord Blood Bank (I-cord, Chabiotech Co., Inc., Seoul, Korea) under approval of IRB of CHA General Hospital, Seoul, Korea. Mononuclear cells (MNCs) were isolated following density gradient separation with Ficoll and CD34+ cells were positively selected directly from MNCs by immunomagnetic cell separation. Female NOD/SCID mice, 8 to 12 weeks of age, were maintained under specific pathogen-free conditions. MNCs (2x107) or CD34+ cells (2x105) derived from UCB were transplanted into NOD/SCID mice by intravenous injection into a lateral tail vein. Two or 4 weeks after transplantation, uterus of mice was recovered and sampled for PCR of human specific primer set (the α-satellite DNA on human chromosome 17) and immunohistochemistry using anti-human nuclear antigen antibody (Chemicon) and peroxidase/DAB staining kit (DAKO). ResultsAt 2 weeks, signal of human specific primer set was not detected in uterus of MNCs and CD34+ cells-transferred mice. However, strong signals were detected in uterus of all of transplanted mice after 4 weeks. Immunohistochemistry revealed human cells originated from MNCs and CD34+ cells in stroma and glandular epithelium of mouse uterus. At 2 weeks, signal of human specific primer set was not detected in uterus of MNCs and CD34+ cells-transferred mice. However, strong signals were detected in uterus of all of transplanted mice after 4 weeks. Immunohistochemistry revealed human cells originated from MNCs and CD34+ cells in stroma and glandular epithelium of mouse uterus. ConclusionThese preliminary finding suggest that human UCB stem cells can incorporate into the uterus by intravenous injection and may sustain. Functional regeneration to endometrial cells will be further investigated. These preliminary finding suggest that human UCB stem cells can incorporate into the uterus by intravenous injection and may sustain. Functional regeneration to endometrial cells will be further investigated.