Experimental transplantation study for possible transformation of bone marrow cells in the mouse placenta.
The aim of the present study is to establish a mouse model of the transplantation of bone marrow cells into the placenta in mid-gestation. The mononuclear fraction of bone marrow cells was isolated by Ficoll gradient centrifugation from the femur bones of C57BL/6 green fluorescent protein (GFP) gene transgenic (Tg) mice. After intraperitoneal injection of pentobarbital sodium, the abdominal cavities of pregnant non-Tg (C57BL/6 or ICR) mice were opened at 9.5 days postcoitum (dpc). The mononuclear fraction of bone marrow cells from Tg mice (3-5 x 10(5)cells/3 microl) was directly injected into the placental portion of the pregnant uterus, at a depth of approximately 3 mm, using a 31-gauge injector. The placenta was sampled at 14.5 dpc. Confocal laser scanning microscopic analysis of the serial sections of the sampled placenta (150-250 sections/placenta) was carried out to detect GFP-positive cells and to assess immunostaining for cytokeratin, CD34, p57(Kip2) and prolactin. Most pregnant mice survived until sampling of the placenta at 14.5-18.5 dpc (88.9% for C57BL6 and 100% for ICR). The survival rate of fetuses from mice in which the placenta was transplanted with GFP-positive bone marrow cells was approximately 50%. A small population (0.154%) of injected bone marrow cells was retained in the placental tissue. Immunohistochemically, cytokeratin, CD34 and p57(Kip2) were positively stained in 0.062%, 4.5% and 2.1% of GFP-positive cells, respectively, while prolactin was not positive in any of the cells examined. GFP-positive bone marrow cells were successfully transplanted to the murine placenta. Future investigations of the specific antigens in bone marrow cells retained in the placenta may enable a better understanding of the local regulation of placental development.
Kakui
K
K
Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Sakyo-ku, Japan.
Itoh
H
H
Sagawa
N
N
Yura
S
S
Takemura
M
M
Kawamura
M
M
Fujii
S
S
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation; D005260:Female; D005865:Gestational Age; D006085:Graft Survival / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007202:Indicators and Reagents / Q000378:metabolism; D008136:Longevity; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D023421:Models, Animal; D009000:Monocytes / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D011247:Pregnancy; D013047:Specific Pathogen-Free Organisms; D018183:Transplantation Chimera; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism*
D015415:Biomarkers; D007202:Indicators and Reagents; D049452:Green Fluorescent Proteins