Cell populations comprising cd31-positive, cd45-negative, cd200-positive mammalian cells, and use thereof
A technology of mammals and cell groups, applied to animal cells, vertebrate cells, cells modified by introducing foreign genetic material, etc., to improve blood supply deficiency, low nutrition, and ischemia
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Embodiment 1
[0107] [Example 1: Identification of mouse liver vascular endothelial stem cells based on cell surface markers]
[0108] Using the SP cell population and the MP cell population obtained from the vascular endothelial cells (CD31-positive CD45-negative cells) of the mouse liver, comprehensively analyzed the genes expressed more in the SP cell population than in the MP cell population, and found out More than 100 genes. From these genes an attempt was made to identify cell surface markers of vascular endothelial stem cells present in about 10% of the SP cell population.
[0109] 1-1 Analysis of surface markers of mouse liver vascular endothelial cells
[0110] (1) Made using animals and cells
[0111] C57BL / 6 mice and C57BL / 6-Tg(CAG-EGFP) mice (commonly known as green mice) were purchased from Japan SLC, Inc. Mice aged 8 to 12 weeks were used in the experiments. The mice were laparotomed under anesthesia, and the liver was removed. Cut the liver into pieces, immerse in a m...
Embodiment 2
[0132] [Example 2: Treatment of hemophilia based on transplantation of CD157-positive CD200-positive vascular endothelial cells]
[0133] Since stem cells can continue to generate terminally differentiated somatic cells while maintaining their undifferentiated nature, transplantation of CD157-positive and CD200-positive cells, which are considered to be vascular endothelial stem cells, can be expected to continue contributing as vascular endothelial cells for a long time. Therefore, it is considered that transplantation of these cells to a patient suffering from a disease caused by dysfunction of vascular endothelial cells may potentially cure the disease. Therefore, it was investigated whether transplantation of normal CD157-positive CD200-positive cells into the liver of hemophilia A model mice could suppress the bleeding tendency of hemophilia A model mice.
[0134] (1) Experimental method
[0135] Hemophilia A model mice (coagulation factor VIII gene-deficient mice) were ...
Embodiment 3
[0142] [Example 3: Liver regeneration based on transplantation of CD157-positive CD200-positive vascular endothelial cells]
[0143] In recent years, it has been reported that molecules secreted by vascular endothelial cells play an important role in maintaining tissue homeostasis or tissue remodeling. For the liver, the maintenance mechanism of liver cells guided by vascular endothelial cells in hepatic sinusoidal vessels has also been elucidated. . Since it is known that CD157-positive CD200-positive vascular endothelial cells contribute significantly to the formation of hepatic sinusoidal vessels, it was investigated whether transplantation of CD157-positive CD200-positive vascular endothelial cells can promote liver regeneration.
[0144] 3-1 Liver regeneration experiment 1
[0145] (1) Experimental method
[0146] 70% partial hepatectomy was performed on C57BL / 6 mice according to conventional methods. According to the method described in 1-1 of Example 1, CD157-positiv...
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