OBJECTIVES: To reprogram human placental fibroblasts (HPFs) into chemically induced epithelioid-like cells (ciEP-Ls) using a combination of small-molecule compounds. METHODS: HPFs cultured under normoxic conditions were identified using immunofluorescence assay, PCR and chromosomal karyotyping. Under hypoxic conditions (37 ℃, 5% O₂), HPFs were cultured in a medium containing small-molecule compounds C6TSEDRVAJZ (CHIR99021, 616452, TTNPB, SAG, EPZ5676, DZNep, Ruxolitinib, VTP50469, Afuresertib, JNK-IN-8, and EZM0414), and the cell morphology was observed daily. The expression levels of epithelial cell markers in the induced cells were detected by immunofluorescence, Western blotting and PCR. Chromosomal karyotyping of the induced cells was performed and the induction efficiency was calculated. RESULTS: Before induction, HPFs showed positive expressions of fibroblast surface markers CD34 and vimentin and were negative for epithelial surface markers. PCR results showed high expressions of fibroblast-specific genes S100A4 and COL1A1 in HPFs with a normal human diploid karyotype. After one day of induction, the HPFs underwent morphological changes from a multinodular spindle shape to a round or polygonal shape, which was morphologically characteristic of ciEP-Ls. On day 4 of induction, the cells exhibited high expressions of the epithelial cell markers E-cadherin and Lin28A. RT-qPCR results also showed that the cells expressed the epithelial markers Smad3, GLi3, PAX8, WT1, KRT19, and KRT18 with significantly down-regulated expressions of all the fibroblast surface markers and a normal human diploid karyotype. The reprogramming efficiency of HPFs into ciEP-Ls ranged from (64.53±2.8)% to (68.10±3.6)%. CONCLUSIONS The small-molecule compound combination C6TSEDRVAJZ is capable of inducing HPFs into ciEP-Ls under hypoxic conditions with a high induction efficiency.
Humans ; Fibroblasts/drug effects* ; Pregnancy ; Female ; Cell Differentiation/drug effects* ; Pyrimidines/pharmacology* ; Placenta/cytology* ; Cells, Cultured ; Pyridines/pharmacology* ; Pyrazoles/pharmacology* ; Epithelial Cells/cytology*

OBJECTIVES: To investigate the impact of prenatal fear stress on placental amino acid transport and emotion and cognition development in offspring rats. METHODS: Thirty pregnant Wistar rats were randomized equally into control and fear stress (induced using an observational foot shock model) groups. In each group, placental and serum samples were collected from 6 dams on gestational day 20, and the remaining rats delivered naturally and the offspring rats were raised under the same conditions until 8 weeks of age. Emotional and cognitive outcomes of the offspring rats were assessed with behavioral tests, and placental structure was examined using HE staining. Bioinformatics analysis was used to identify differentially expressed placental transporter genes under fear stress. The expressions of system A and system L amino acid transporters, along with other specialized transporters, were detected using qRT-PCR and Western blotting. Fetal serum amino acid concentrations were determined by HPLC. The correlations between fetal amino acid levels and behavioral outcomes of the offspring rats were analyzed. RESULTS: The dams with fear stress showed reduced open-field activity and increased freezing behavior with significantly decreased placental weight, fetal weight, and fetal-to-placental ratio. Bioinformatics analysis revealed 28 differentially expressed transporter genes involved mainly in amino acid transport. In the fear stress group, fetal serum amino acid levels were significantly lowered and Slc38a1, Slc43a1, Slc43a2, Slc7a8, Slc6a6, Slc1a1 and Slc6a9 mRNA and protein expressions were all downregulated. The offspring rats in fear stress group exhibited decreased novel object preference and spontaneous alternation with reduced open arm exploration and increased immobility in emotional tests. Lower early-life amino acid levels was found to correlate with impaired adult cognition. CONCLUSIONS Prenatal fear stress in rats impairs placental amino acid transporter expression and reduces fetal serum amino acid levels, potentially contributing to long-term cognitive deficits in the offspring rats.
Animals ; Female ; Pregnancy ; Placenta/metabolism* ; Fear ; Rats ; Rats, Wistar ; Cognition ; Prenatal Exposure Delayed Effects ; Stress, Psychological ; Amino Acids/blood* ; Amino Acid Transport Systems/metabolism*

Trophoblast cells serve as the foundation for placental development. We analyzed published multiomics sequencing data and found that trophoblast cells highly expressed RRS1 compared to primitive endoderm and epiblast. We used HTR-8/SVneo cells for further investigation, and Western blot and immunofluorescence staining confirmed that HTR-8/SVneo cells highly expressed RRS1. RRS1 was successfully knocked down in HTR-8/SVneo cells using siRNA. Using IncuCyte S3 live-cell analysis system based on continuous live-cell imaging and real-time data, we observed that proliferation, migration, and invasion abilities were all significantly decreased in RRS1-knockdown cells. RNA-seq revealed that knockdown of RRS1 affected the gene transcription, and upregulated pathways in extracellular matrix organization, DNA damage response, and intrinsic apoptotic signaling, downregulated pathways in embryo implantation, trophoblast cell migration, and wound healing. Differentially expressed genes were enriched in diseases related to placental development. Consistent with these findings, human chorionic villus samples collected from spontaneous abortion cases exhibited significantly reduced RRS1 expression compared to normal controls. Our results highlight the functional importance of RRS1 in human trophoblasts and suggest that its deficiency contributes to early pregnancy loss.
Humans ; Trophoblasts/physiology* ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Female ; Pregnancy ; Abortion, Spontaneous/metabolism* ; Cell Line ; Placentation/genetics*

A 40-year-old, gravida 3 para 2 (1-1-0-2), previous primary cesarean section for nonreassuring fetal status, presented at a tertiary hospital for confirmation of cesarean scar pregnancy (CSP). Transvaginal ultrasound confirmed a CSP at 8 2/7 weeks age of gestation with good embryonic cardiac activity, raising concern for early placenta accreta spectrum. A multidisciplinary team composed of an obstetrician, advanced pelvic surgeon, urologist, and anesthesiologist managed the patient. The patient underwent total abdominal hysterectomy with bilateral salpingectomy, as the patient has a completed family size. Before the procedure, she was given cefuroxime as prophylactic antibiotic. Intraoperatively, there were dense adhesions between the posterior bladder wall and the previous cesarean section scar. Inadvertent injury to the bladder wall was incurred during adhesiolysis. Cystorrhaphy was done by a urologist, while the rest of the surgery was unremarkable, with a 450 ml estimated blood loss. The postoperative course was unremarkable. Bladder rest was achieved by maintaining an indwelling Foley catheter, which remained in place upon discharge on postoperative day 3 and was continued for 7 days thereafter. At follow-up, a successful voiding trial was conducted, confirming the return of normal bladder function.

Human ; Female ; Adult: 25-44 Yrs Old ; Cesarean Section ; Salpingectomy ; Hysterectomy ; Fetal Distress ; Placenta Accreta ; Cefuroxime ; Catheters ; Cicatrix

Objective To explore the alterations in macrophage polarization and the expression of decorin (DCN) protein in the decidua of patients with missed abortion (MA), as well as to elucidate the regulatory effect of DCN on macrophage polarization. Methods Flow cytometry was employed to assess the polarization ratio of decidual macrophages in MA, recurrent spontaneous abortion (RSA) and normal pregnancy (NP); The expression and localization of DCN and hypoxia-inducible factor 1α (HIF-1α) in decidua and villi were assessed by immunohistochemistry staining, while their protein levels were measured by Western blot. Primary trophoblasts and peripheral blood mononuclear cell (PBMC)-derived macrophages were isolated and cultured. ELISA was conducted to quantify DCN levels in the culture supernatant of primary trophoblast and PBMC-derived macrophages. Additionally, flow cytometry was applied to evaluate the polarization ratio of PBMC-derived macrophages. Immunofluorescence cytochemical staining was conducted to examine HIF-1α expression in macrophages. Western blot was performed to detect the expression of proteins related to the gene associated with retinoid-IFN-induced mortality 19 (GRIM-19)/signal transducer and activator of transcription 3 (STAT3)/HIF-1α signaling pathway in macrophages. Results The polarization ratio of M1 macrophages in the decidua of abortion patients was significantly higher than that of NP, whereas the ratio of M2 macrophages was significantly lower. The expression of DCN and HIF-1α protein were significantly evaluated in abortion patients compared to NP. The supernatant DCN content and HIF-1α protein expression of primary trophoblast and PBMC-derived macrophages cultured under 10 mL/L O₂ for 24 hours were markedly increased compared to cells treated with 210 mL/L O₂. Compared with the PBS group, the proportion of M1 macrophage and GRIM-19 protein expression were significantly reduced in the DCN group, while phosphorylated STAT3 (p-STAT3) and HIF-1α protein expression were significantly increased. Conclusion The expression of DCN in decidua and villi of MA is higher than that of NP. DCN exhibits an inhibitory effect on the M1 polarization of PBMCs-derived macrophages, which is likely mediated through the GRIM-19/STAT3/HIF-1α signaling pathway.
Humans ; Female ; Decidua/metabolism* ; Macrophages/cytology* ; Decorin/genetics* ; Pregnancy ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Adult ; Leukocytes, Mononuclear/cytology* ; Abortion, Missed/genetics* ; STAT3 Transcription Factor/metabolism* ; Cells, Cultured ; Young Adult

Severe combined immunodeficiency (SCID) represents a group of genetically heterogeneous disorders characterized by mutations that lead to profound defects in both humoral and cellular immunity. Transplacental maternal engraftment (TME) is a frequently observed complication in SCID. While most cases of SCID with TME exhibit no substantial impact on disease progression, a subset of patients may encounter diagnostic delays or therapeutic challenges due to TME interference. Furthermore, TME may predispose these individuals to graft-versus-host disease (GVHD) prior to hematopoietic stem cell transplantation, thereby increasing diagnostic complexity and treatment risks. This review systematically examines the etiology and clinical manifestations of SCID associated with TME, analyzes its implications for disease management, and evaluates current detection methodologies. The synthesized evidence provides a theoretical foundation for future research and offers potential insights into the clinical diagnosis and management of SCID associated with TME.
Humans ; Severe Combined Immunodeficiency/diagnosis* ; Pregnancy ; Female ; Hematopoietic Stem Cell Transplantation/adverse effects* ; Maternal-Fetal Exchange/immunology* ; Graft vs Host Disease/etiology* ; Animals ; Placenta/immunology*

OBJECTIVE: To investigate the role of extracellular vesicles (EVs) derived from placental tissue and placental mesenchymal stem cells in supporting the growth and function of adult hematopoietic stem and progenitor cells (HSPCs), so as to optimize their culture system. METHODS: EVs were isolated from mouse placental tissue (PL-EV) and placental mesenchymal stem cells (PL-MSC-EV). These EVs were co-cultured with 3 000 adult bone marrow LKS⁺ (lineage^- c-Kit⁺ Sca-1⁺ ) cells for 72 hours at concentrations of 0, 1, 10, 50, 100, and 200 μg/ml. The proportion and absolute count of LKS⁺ cells after co-culture were analyzed by flow cytometry, while their self-renewal and multi-lineage differentiation potential were evaluated using colony-forming unit (CFU) assays. RESULTS: Compared to the blank control group, the proportion of LKS⁺ cells were significantly increased in PL-EV groups at concentrations ≥10 μg/ml after 72 hours of co-culture. Notably, LKS⁺ cells co-cultured at the concentration of 10 μg/ml exhibited the highest absolute count (899±171) and the highest proportion of LT-HSCs (LKS⁺ CD135^- CD34^-) (0.67%±0.07%). In the PL-MSC-EV co-culture system, the absolute count of LKS⁺ cells peaked at the concentration of 1 μg/ml (1011±99 cells), though the proportion of LT-HSCs was relatively low (0.15%±0.05%). The comparison between these two culture systems revealed that PL-EV at 10 μg/ml and PL-MSC-EV at 1 μg/ml displayed the most pronounced effects on LKS⁺ cell proliferation, but with no significant difference between them. CFU assays showed that, in the PL-EV culture system, the number of LKS⁺ colony formed in 1 and 10 μg/ml groups was not significantly different compared with the blank control group. In contrast, in the PL-MSC-EV system, the highest LKS⁺ colony-forming capacity was observed when co-cultured with 1 μg/ml PL-MSC-EV, while a significant reduction was noted at concentrations above 10 μg/ml. CONCLUSION PL-EV and PL-MSC-EV effectively support the growth and function of HSPCs. And PL-MSC-EV exhibits a superior efficacy in preserving the stemness of LKS⁺ cells, thus suggesting its potential for optimizing culture systems of HSPCs.
Mesenchymal Stem Cells/cytology* ; Extracellular Vesicles ; Placenta/cytology* ; Female ; Animals ; Mice ; Pregnancy ; Hematopoietic Stem Cells/cytology* ; Coculture Techniques ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured

Retained products of conception (RPOC) represent a common pregnancy-related condition that may lead to complications such as abnormal uterine bleeding, infection, secondary arteriovenous fistula, intrauterine adhesions, and infertility. Currently, the main clinical treatments for RPOC include surgical intervention, medical therapy, and expectant management, sometimes supplemented by high-intensity focused ultrasound or uterine artery embolization when necessary. However, no standardized treatment guidelines exist. Medical and expectant management may help some patients avoid or reduce the need for surgery, though these approaches often involve a prolonged disease course. While surgery yields rapid results, patients with large lesions may require multiple procedures, increasing the risk of endometrial damage and intrauterine adhesions. There is still a lack of robust evidence-based guidance for selecting the optimal or individualized treatment approach. This review explores recent advances in the management of RPOC, with an emphasis on strategies that effectively preserve the endometrium, safeguard fertility, and support more precise, minimally invasive, and efficient personalized treatment.
Humans ; Female ; Pregnancy ; Placenta, Retained/surgery* ; Uterine Artery Embolization/methods* ; Tissue Adhesions ; Endometrium ; High-Intensity Focused Ultrasound Ablation

Preeclampsia (PE) is a severe gestational disorder characterized by hypertension and proteinuria, with a subset of cases exhibiting an immune-driven phenotype marked by placental overexpression of proinflammatory cytokines and chronic inflammatory damage, profoundly impacting fetal development. To elucidate the pathophysiology of this PE subtype, we established an inflammation-driven PE mouse model via lipopolysaccharide (LPS) intraperitoneal injection, systematically evaluating histopathological changes in maternal heart, liver, lung, kidney, and placenta, and integrating transcriptomic profiling to uncover molecular mechanisms. LPS administration robustly induced maternal hypertension and proteinuria, hallmarks of PE, without significantly altering organ or fetal weights. Histological analyses revealed pronounced inflammatory damage in the maternal lung, kidney, and placenta, with the lung exhibiting the most severe pathology, characterized by inflammatory cell infiltration, alveolar wall thickening, and interstitial edema-challenging the conventional focus on placental and renal primacy in PE. Placental labyrinth and junctional zones displayed extensive structural disruption and necrosis, indicating functional impairment. Transcriptomic analysis identified 27 inflammation-related genes consistently upregulated across tissues, with protein-protein interaction networks pinpointing Il1β, Il6, Ccl5, Ccl2, Cxcl10, Tlr2, and Icam1 as hub genes. Quantitative PCR validation confirmed Tlr2 as a central regulator, evidenced by significant upregulation of Tlr2 in lung, kidney, and placenta of LPS-induced PE mice, while Cxcl10 exhibited placenta-specific upregulation, suggesting a synergistic inflammatory axis in placental pathology. These findings highlight the lung as a critical, yet underappreciated, target in inflammation-driven PE, reframe the multi-organ inflammatory landscape of the disease, and nominate Tlr2 and Cxcl10 as potential diagnostic biomarkers and therapeutic targets, offering new avenues for precision intervention in PE.
Animals ; Female ; Pregnancy ; Mice ; Pre-Eclampsia/genetics* ; Inflammation ; Lipopolysaccharides/adverse effects* ; Disease Models, Animal ; Transcriptome ; Placenta/pathology* ; Phenotype

The onset of pregnancy is marked by the formation of a zygote, while the culmination of gestation is manifested by the delivery of a fetus. Meanwhile, a successful pregnancy entails a meticulously coordinated sequence of events from embryo implantation to sustained decidualization of the uterus to placental development and childbirth. The decidual reaction, a pivotal process occurring within the endometrium during pregnancy, is finely regulated by sex steroids and cytokines. Notably, fibroblast growth factors (FGFs), particularly FGF2, play a critical role in this physiological cascade. Dysregulated FGF expression may trigger inadequate decidualization, precipitating a spectrum of adverse pregnancy outcomes, including preeclampsia, recurrent implantation failure, and miscarriage. Furthermore, the human decidua, distinct from most mammalian species and similar to great apes, undergoes regular cycles of formation and shedding, independent of the presence of the embryo in the endometrium. This process is also tightly controlled by various FGFs. In this review, we comprehensively compare diverse research decidualization models, delineate the trend of endometrial FGFs during the menstrual cycle, and provide a synopsis of endometrial diseases triggered by FGF dysregulation.
Humans ; Female ; Pregnancy ; Decidua/physiology* ; Animals ; Endometrium/physiology* ; Fibroblast Growth Factors/metabolism* ; Embryo Implantation ; Menstrual Cycle/physiology*