Generation and characterization of human Fetal membrane and Decidual cell lines for reproductive biology experiments†.

Human fetal membrane and maternal decidua parietalis form one of the major feto-maternal interfaces during pregnancy. Studies on this feto-maternal interface is limited as several investigators have limited access to the placenta, and experience difficulties to isolate and maintain primary cells. Many cell lines that are currently available do not have the characteristics or properties of their primary cells of origin. Therefore, we created, characterized the immortalized cells from primary isolates from fetal membrane-derived amnion epithelial cells, amnion and chorion mesenchymal cells, chorion trophoblast cells and maternal decidua parietalis cells. Primary cells were isolated from a healthy full-term, not in labor placenta. Primary cells were immortalized using either a HPV16E6E7 retroviral or a SV40T lentiviral system. The immortalized cells were characterized for the morphology, cell type-specific markers, and cell signalling pathway activation. Genomic stability of these cells was tested using RNA seq, karyotyping, and short tandem repeats DNA analysis. Immortalized cells show their characteristic morphology, and express respective epithelial, mesenchymal and decidual markers similar to that of primary cells. Gene expression of immortalized and primary cells were highly correlated (R = 0.798 to R = 0.974). Short tandem repeats DNA analysis showed in the late passage number (>P30) of cell lines matched 84-100% to the early passage number (

Postnatal depletion of maternal cells biases T lymphocytes and natural killer cells' profiles toward early activation in the spleen.

The maternal cells transferred into the fetus during gestation persist long after birth in the progeny. These maternal cells have been hypothesized to promote the maturation of the fetal immune system in utero but there are still significant gaps in our knowledge of their potential roles after birth. To provide insights into these maternal cells' postnatal functional roles, we set up a transgenic mouse model to specifically eliminate maternal cells in the neonates by diphtheria toxin injection and confirmed significant depletion in the spleens. We then performed immunophenotyping of the spleens of two-week-old pups by mass cytometry to pinpoint the immune profile differences driven by the depletion of maternal cells in early postnatal life. We observed a heightened expression of markers related to activation and maturation in some natural killer and T cell populations. We hypothesize these results to indicate a potential postnatal regulation of lymphocytic responses by maternal cells. Together, our findings highlight an immunological influence of maternal microchimeric cells postnatally, possibly protecting against adverse hypersensitivity reactions of the neonate at a crucial time of new encounters with self and environmental antigens.

Detection of Maternal Cells in Human Umbilical Cord Blood Using PCR and Fluorescence In Situ Hybridization / 대한혈액학회지

BACKGROUND: Cord blood is a practical source of hematopoietic stem cells for transplantation. However, there are concerns that cord blood might be contaminated with maternal cells that could lead to graft-versus-host disease. MATERIAL AND METHODS: In the present study, we used the polymerase chain reaction (PCR) amplification of two minisatellite sequences (YNZ 22 and 33.6) and fluorescence in situ hybridization (FISH), to ascertain the presence of maternal cells in the cord blood. RESULTS: The results were as follows; 1) Maternal-specific allele was present in 1 of the 16 cord bloods (6.25%) by PCR. 2) We determined the sensitivity of this methods for detecting a maternal specific allele in the cord blood sample, which demonstrated as 0.2% (YNZ 22 and 33.6). 3) Maternal cells were found in 11 of the 20 cord bloods (55%), and 35 XX maternal cells were observed in 19,138 nuclei in the cord blood by FISH. 4) The range of maternal cells in the 11 positives was 0.1~8.6% and median value of maternal cells was 0.2%. CONCLUSION: These results suggest that maternal cells are very rarely present in the cord blood collected at birth. However, although small, this amount of cells may result in GVHD in a susceptible recipient. These methods we used allow the detection of maternal cells within cord blood from 104 nucleated cells by FISH but it was difficult to detect maternal cells within female cord blood by PCR, then it needs the use of a more reliable tool in a cord blood banking perspective.