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Comprehensive analysis of leukocytes, vascularization and matrix metalloproteinases in human menstrual xenograft model.

Guo Y, He B, Xu X, Wang J - PLoS ONE (2011)

Bottom Line: Human CD45+ cells showed a peak of increase 28 days post-transplantation.Three days after progesterone withdrawal, mouse CD45+ cells increased rapidly in number and were significantly greater than human CD45+ cell counts.After progesterone withdrawal, the expression levels of matrix metalloproteinases (MMP) 1, 2, and 9 were increased.

View Article: PubMed Central - PubMed

Affiliation: Graduate School, Peking Union Medical College, Beijing, People's Republic of China.

ABSTRACT
In our previous study, menstrual-like changes in mouse were provoked through the pharmacologic withdrawal of progesterone with mifepristone following induction of decidualization. However, mouse is not a natural menstruation animal, and the menstruation model using external stimuli may not truly reflect the occurrence and development of the human menstrual process. Therefore, we established a model of menstruation based on human endometrial xenotransplantation. In this model, human endometrial tissues were transplanted subcutaneously into SCID mice that were ovarectomized and supplemented with estrogen and progestogen by silastic implants with a scheme imitating the endocrinological milieu of human menstrual cycle. Morphology, hormone levels, and expression of vimentin and cytokeratin markers were evaluated to confirm the menstrual-like changes in this model. With 28 days of hormone treatment, transplanted human endometrium survived and underwent proliferation, differentiation and disintegration, similar to human endometrium in vivo. Human CD45+ cells showed a peak of increase 28 days post-transplantation. Three days after progesterone withdrawal, mouse CD45+ cells increased rapidly in number and were significantly greater than human CD45+ cell counts. Mouse CD31+ blood vascular-like structures were detected in both transplanted and host tissues. After progesterone withdrawal, the expression levels of matrix metalloproteinases (MMP) 1, 2, and 9 were increased. In summary, we successfully established a human endometrial xenotransplantation model in SCID mice, based on the results of menstrual-like changes in which MMP-1, 2 and 9 are involved. We showed that leukocytes are originated from in situ proliferation in human xenografts and involved in the occurrence of menstruation. This model will help to further understand the occurrence, growth, and differentiation of the endometrium and the underlying mechanisms of menstruation.

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Immunofluorescence for human vimentin and cytokeratin after transplantation.The harvested tissues were detected by dual immunofluorescence for human vimentin and cytokeratin 28 days after transplantation. Stromal cells were specifically stained with vimentin (green), while epithelial cells were specifically stained with cytokeratin (red). Nuclei were visualized by DAPI (blue). Bars: 100 µm. Figure is from serial thin sections. Original magnification: 400×.
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pone-0016840-g004: Immunofluorescence for human vimentin and cytokeratin after transplantation.The harvested tissues were detected by dual immunofluorescence for human vimentin and cytokeratin 28 days after transplantation. Stromal cells were specifically stained with vimentin (green), while epithelial cells were specifically stained with cytokeratin (red). Nuclei were visualized by DAPI (blue). Bars: 100 µm. Figure is from serial thin sections. Original magnification: 400×.

Mentions: The anti-human vimentin and cytokeratin antibodies used in our study do not cross-react with mouse tissues, and can specifically label transplanted human tissues in mice. The green fluorescence, red fluorescence and blue fluorescence showed human stromal cells, human epithelial cells, and DAPI (Sigma, St. Louis, Missouri, USA)-labeled nuclei, respectively. As shown in Fig. 4, the areas outside the regions of green and red fluorescence were mouse tissues, while the green-fluorescence-positive and -negative regions were clearly demarcated. Fibrous connective tissues around the xenotransplanted tissues were green-fluorescence negative, indicating that they were mouse tissues, rather than human endometrial tissues.


Comprehensive analysis of leukocytes, vascularization and matrix metalloproteinases in human menstrual xenograft model.

Guo Y, He B, Xu X, Wang J - PLoS ONE (2011)

Immunofluorescence for human vimentin and cytokeratin after transplantation.The harvested tissues were detected by dual immunofluorescence for human vimentin and cytokeratin 28 days after transplantation. Stromal cells were specifically stained with vimentin (green), while epithelial cells were specifically stained with cytokeratin (red). Nuclei were visualized by DAPI (blue). Bars: 100 µm. Figure is from serial thin sections. Original magnification: 400×.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3040756&req=5

pone-0016840-g004: Immunofluorescence for human vimentin and cytokeratin after transplantation.The harvested tissues were detected by dual immunofluorescence for human vimentin and cytokeratin 28 days after transplantation. Stromal cells were specifically stained with vimentin (green), while epithelial cells were specifically stained with cytokeratin (red). Nuclei were visualized by DAPI (blue). Bars: 100 µm. Figure is from serial thin sections. Original magnification: 400×.
Mentions: The anti-human vimentin and cytokeratin antibodies used in our study do not cross-react with mouse tissues, and can specifically label transplanted human tissues in mice. The green fluorescence, red fluorescence and blue fluorescence showed human stromal cells, human epithelial cells, and DAPI (Sigma, St. Louis, Missouri, USA)-labeled nuclei, respectively. As shown in Fig. 4, the areas outside the regions of green and red fluorescence were mouse tissues, while the green-fluorescence-positive and -negative regions were clearly demarcated. Fibrous connective tissues around the xenotransplanted tissues were green-fluorescence negative, indicating that they were mouse tissues, rather than human endometrial tissues.

Bottom Line: Human CD45+ cells showed a peak of increase 28 days post-transplantation.Three days after progesterone withdrawal, mouse CD45+ cells increased rapidly in number and were significantly greater than human CD45+ cell counts.After progesterone withdrawal, the expression levels of matrix metalloproteinases (MMP) 1, 2, and 9 were increased.

View Article: PubMed Central - PubMed

Affiliation: Graduate School, Peking Union Medical College, Beijing, People's Republic of China.

ABSTRACT
In our previous study, menstrual-like changes in mouse were provoked through the pharmacologic withdrawal of progesterone with mifepristone following induction of decidualization. However, mouse is not a natural menstruation animal, and the menstruation model using external stimuli may not truly reflect the occurrence and development of the human menstrual process. Therefore, we established a model of menstruation based on human endometrial xenotransplantation. In this model, human endometrial tissues were transplanted subcutaneously into SCID mice that were ovarectomized and supplemented with estrogen and progestogen by silastic implants with a scheme imitating the endocrinological milieu of human menstrual cycle. Morphology, hormone levels, and expression of vimentin and cytokeratin markers were evaluated to confirm the menstrual-like changes in this model. With 28 days of hormone treatment, transplanted human endometrium survived and underwent proliferation, differentiation and disintegration, similar to human endometrium in vivo. Human CD45+ cells showed a peak of increase 28 days post-transplantation. Three days after progesterone withdrawal, mouse CD45+ cells increased rapidly in number and were significantly greater than human CD45+ cell counts. Mouse CD31+ blood vascular-like structures were detected in both transplanted and host tissues. After progesterone withdrawal, the expression levels of matrix metalloproteinases (MMP) 1, 2, and 9 were increased. In summary, we successfully established a human endometrial xenotransplantation model in SCID mice, based on the results of menstrual-like changes in which MMP-1, 2 and 9 are involved. We showed that leukocytes are originated from in situ proliferation in human xenografts and involved in the occurrence of menstruation. This model will help to further understand the occurrence, growth, and differentiation of the endometrium and the underlying mechanisms of menstruation.

Show MeSH
Related in: MedlinePlus