primary endometrial stromal fibroblasts  (ATCC)

Journal: bioRxiv

Article Title: Mechanical Cues Regulate Estrogen and Progesterone-Induced Nascent ECM Deposition by Human Endometrial Stromal Cells

doi: 10.1101/2025.10.14.682403

Figure Lengend Snippet: a Schematic illustrating the female reproductive system including the uterus with the endometrium and myometrium, the ovary and fallopian tubes. Inset shows endometrial stromal cells embedded in extracellular matrix (ECM) containing a variety of proteins (e.g., laminin, collagens and fibronectin). b Schematic illustrating the relative levels of estrogen and progesterone during the menstrual cycle. c Schematic illustrating the endometrial tissue during the proliferative and remodeling phase of the menstrual cycle.

Article Snippet: Primary endometrial stromal cells (PCS-460-010, ATCC; 25-year-old donor) were cultured in fibroblast growth media, containing 10% fetal bovine serum (PCS-201-030 and PCS-201-041, ATCC), and passaged twice prior cryopreservation.

Techniques:

Journal: bioRxiv

Article Title: Mechanical Cues Regulate Estrogen and Progesterone-Induced Nascent ECM Deposition by Human Endometrial Stromal Cells

doi: 10.1101/2025.10.14.682403

Figure Lengend Snippet: a Schematic illustrating the structure of progesterone, estrogen, and 3’,5’-cyclic adenosine monophosphate (cAMP). b Timeline of endometrial stromal cell seeding on glass (-24 hours) and culture for 24 hours to reach confluency before an additional culture up to 96 hours without hormones (Ctrl) or with hormones/cAMP (+Hrm). Cells were fixed at 0, 24, 48, 72, and 96 hours for image analysis. c Schematic illustrating the change in endometrial stromal cell morphology in response to hormonal treatment leading to the secretion of hormones, such as prolactin. d Representative fluorescent images of Phalloidin (F-actin) and Hoechst (nucleus) of endometrial stromal cells cultured for 0, 24, and 48 hours without hormones (Ctrl, scale bars = 100 µm, insets = 55 µm). e Quantification of size and number of nuclei of endometrial stromal cells cultured for 0, 24, and 48 hours in control media (n = 3 replicates, 5 images per replicate from one representative experiment, **p≤0.001, **** p≤0.0001, ns = not significant by one-way ANOVA). f Representative fluorescent images of Phalloidin (F-actin) and Hoechst (nucleus) of endometrial stromal cells cultured for 0, 24, and 48 hours with hormones (+Hrm, scale bars = 100µm, inset = 55µm). g Quantification of size and number of nuclei of endometrial stromal cells cultured for 0, 24, and 48 hours in hormone media (n = 3 replicates, 5 images per replicate from one representative experiment, **p≤0.001, **** p≤0.0001, ns = not significant by one-way ANOVA).

Article Snippet: Primary endometrial stromal cells (PCS-460-010, ATCC; 25-year-old donor) were cultured in fibroblast growth media, containing 10% fetal bovine serum (PCS-201-030 and PCS-201-041, ATCC), and passaged twice prior cryopreservation.

Techniques: Cell Culture, Control

Journal: bioRxiv

Article Title: Mechanical Cues Regulate Estrogen and Progesterone-Induced Nascent ECM Deposition by Human Endometrial Stromal Cells

doi: 10.1101/2025.10.14.682403

Figure Lengend Snippet: a Timeline of endometrial stromal cell seeding: culture for 24 hours to attach to the substrate before additional culture for 48 hours without hormones (Ctrl) or with hormones/cAMP (+Hrm) prior harvesting for image analysis. b Schematic illustrating norbornene-modified hyaluronic acid (NorHA) hydrogels with different elastic moduli using 1.29 (low) or 3.24 mM (high) dithiol. c Quantification of compressive Young’s modulus of 1.29 mM (low) and 3.24 mM (high) DTT crosslinked NorHA hydrogels (n = 3 hydrogels, ***p=0.0001 by Student’s t -test). d Representative fluorescent images of endometrial stromal cells cultured for 48 hours atop 5 kPa and 15 kPa hydrogels without hormones (Ctrl, scale bars = 100µm, inset = 55µm). e Quantification of number of nuclei and area of nascent ECM deposition, normalized to number of nuclei, of endometrial stromal cells cultured for 48 hours atop 5 kP and 15 kPa hydrogels (5kPa: n = 48 images from 4 independent experiments, 15 kPa: n = 40 images from 3 independent experiments, *p≤0.05, ns = not significant by Students t -test). f Representative fluorescent images and quantification of number of nuclei and area of nascent ECM, normalized to number of nuclei, deposited by endometrial stromal cells cultured for 48 hours atop 5 kPa hydrogels with hormones (+Hrm, scale bars = 100 µm, insets = 55 µm, n = 48 images from 4 independent experiments, ****p≤0.0001 by Students t -test). g Representative fluorescent images and quantification of number of nuclei and area of nascent ECM, normalized to number of nuclei, deposited by endometrial stromal cells cultured for 48 hours atop 15 kPa hydrogels with hormones (+Hrm, scale bars = 100 µm, insets = 55 µm, n = 38 images from 3 independent experiments, *p ≤ 0.05 by Students t -test).

Article Snippet: Primary endometrial stromal cells (PCS-460-010, ATCC; 25-year-old donor) were cultured in fibroblast growth media, containing 10% fetal bovine serum (PCS-201-030 and PCS-201-041, ATCC), and passaged twice prior cryopreservation.

Techniques: Modification, Cell Culture

Journal: bioRxiv

Article Title: Mechanical Cues Regulate Estrogen and Progesterone-Induced Nascent ECM Deposition by Human Endometrial Stromal Cells

doi: 10.1101/2025.10.14.682403

Figure Lengend Snippet: a Representative fluorescent images of cells seeded on 5 kPa gels without (Ctrl) and with hormones (+Hrm), stained with Hoechst (nuclei), Phalloidin (F-actin), DBCO-488 (nascent ECM), and anti-Fibronectin antibody (scale bars = 100 µm, insets = 55 µm). b Quantification of area of fibronectin deposited normalized to number of endometrial stromal cells cultured on 5 kPa hydrogels for 48 hours with and without hormones (5 kPa gels: n = 28 images from 3 independent experiments, ****p≤0.0001 by Student’s t -test). c Representative fluorescent images of cells seeded on 15 kPa gels without and with hormones, stained with Hoechst, Phalloidin, DBCO-488, and anti-Fibronectin antibody (scale bars = 100 µm, insets = 55 µm). d Quantification of area of fibronectin deposited normalized to number of endometrial stromal cells cultured on 15 kPa hydrogels for 48 hours with and without hormones (15 kPa gels: n = 19 images from 2 independent experiments, **p ≤ 0.01 by Student’s t -test). e Representative fluorescent images of cells seeded on glass coverslips without and with hormones, stained with Hoechst, Phalloidin, DBCO-488, and anti-Fibronectin antibody (scale bars = 100 µm, insets = 55 µm). f Quantification of area of fibronectin deposited normalized to number of endometrial stromal cells cultured on glass coverslips for 48 hours with and without hormones (glass: n = 15 images from one representative experiment, ns = not significant by Student’s t -test).

Article Snippet: Primary endometrial stromal cells (PCS-460-010, ATCC; 25-year-old donor) were cultured in fibroblast growth media, containing 10% fetal bovine serum (PCS-201-030 and PCS-201-041, ATCC), and passaged twice prior cryopreservation.

Techniques: Staining, Cell Culture

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: Dkk1 promoted autophagy of endometrial stromal cells. ( A ) After silencing DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of DKK1,LC3-II, ATG7 and P62 proteins. ( B )After knock out of DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of DKK1,LC3-II, ATG7 and P62 proteins. ( C ) After ectopic expression of DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of DKK1,LC3-II, ATG7 and P62 proteins. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively

Article Snippet: Primary human endometrial stromal cells (PHESC) were purchased from Procell Life Science&Technology Co., Ltd. (Cat NO.: CP-H233).

Techniques: Western Blot, Expressing, Knock-Out

Journal: Journal of Translational Medicine

Article Title: DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition

doi: 10.1186/s12967-024-05402-5

Figure Lengend Snippet: DKK1 regulates Wnt/β-catenin and PI3K/AKT/mTOR pathways. ( A ) After knock out of DKK1 in primary endometrial stromal cells, next-generation sequencing was used to detect transcriptome differences between KO group and WT group. Figure ( A ) is a heat map. Figure ( B ) is a volcanic map. ( C ) KEGG signaling pathway analysis of differentially expressed genes. ( D ) After silencing Wnt5a in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of Wnt5a, LC3-II, ATG7 and P62 proteins. ( E ) After silencing TCF4 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of TCF4, LC3-II, ATG7 and P62 proteins. ( F ) After knock out of DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of WNT5A, β-catenin, p-akt, p38 and ATF4 proteins. ( G ) After ectopic expression of DKK1 in primary endometrial stromal cells (PESC) and endometrial stromal cells immortalized cell lines (T-HESCS), western blot analysis was performed to detect the expression of WNT5A, β-catenin, p-akt, p38 and ATF4 proteins. Error bars represent the standard error. The symbols * and ** indicate p < 0.05 and 0.01, respectively

Article Snippet: Primary human endometrial stromal cells (PHESC) were purchased from Procell Life Science&Technology Co., Ltd. (Cat NO.: CP-H233).

Techniques: Knock-Out, Next-Generation Sequencing, Western Blot, Expressing

Journal: Cancer Management and Research

Article Title: Exosomal miR-499a-5p Inhibits Endometrial Cancer Growth and Metastasis via Targeting VAV3

doi: 10.2147/CMAR.S283747

Figure Lengend Snippet: Gene expression profile in endometrial cancer. ( A ) Hierarchical clustering image of different expressed miRNAs between endometrial cancer tissues and adjacent normal tissues from patients. ( B ) Venn diagram was generated to exhibit putative target gene of miR-499 from prediction database, including TargetScan, miRDB, and miRWalk. ( C ) miR-499 and VAV3 expression in endometrial cancer cells and normal endometrium cells (n=3 each group). Data are presented as the means±SD. *** P <0.0001.

Article Snippet: Human normal endometrial stromal cells (SHT290) and human endometrial cancer cell lines, including Ishikawa, HEC1A, and AN3CA, were purchased from American Type Culture Collection (ATCC, VA, USA).

Techniques: Gene Expression, Generated, Expressing