A construct of adipose-derived mesenchymal stem cells-laden collagen scaffold for fertility restoration by inhibiting fibrosis in a rat model of endometrial injury.

Severe endometrium damage causes pathological conditions such as thin endometrium and intrauterine adhesion, resulting in uterine factor infertility. Mesenchymal stem cell (MSC) therapy is a promising strategy in endometrial repair; yet, exogenous MSCs still raise concerns for safety and ethical issues. Human adipose-derived mesenchymal stem cells (ADMSCs) residing in adipose tissue have high translational potentials due to their autologous origin. To harness the high translation potentials of ADMSC in clinical endometrium regeneration, here we constructed an ADMSCs composited porous scaffold (CS/ADMSC) and evaluated its effectiveness on endometrial regeneration in a rat endometrium-injury model. We found that CS/ADMSC intrauterine implantation (i) promoted endometrial thickness and gland number, (ii) enhanced tissue angiogenesis, (iii) reduced fibrosis and (iv) restored fertility. We ascertained the pro-proliferation, pro-angiogenesis, immunomodulating and anti-fibrotic effects of CS/ADMSC in vitro and revealed that the CS/ADMSC influenced extracellular matrix composition and organization by a transcriptomic analysis. Our results demonstrated the effectiveness of CS/ADMSC for endometrial regeneration and provided solid proof for our future clinical study.

Identification of cells with colony-forming activity, self-renewal capacity, and multipotency in ovarian endometriosis.

Endometriosis, the growth of endometrial tissue outside the uterine cavity, is a common gynecological disorder affecting 10% to 15% of women in their reproductive years. Retrograde menstrual shedding containing endometrial stem/progenitor cells has been postulated to be involved in its pathogenesis. In this study, we identified putative endometriotic stem/progenitor cells by their colony-forming potential, self-renewal capacity, and multipotency. Purified epithelial and stromal cells isolated from ovarian endometriotic cysts formed large and small colony-forming units (CFUs) in clonogenic assay. The colony-forming activity of epithelial and stromal cells was found to differ greatly between autologous endometrium and ovarian endometrioma samples. The large CFUs could propagate more than the small CFUs. The endometriotic epithelial small CFUs expressed epithelial markers (epithelial cell adhesion molecule, cytokeratin, and α6 integrin); only occasional large CFUs expressed α6 integrin. Aside from the expression of fibroblast markers, stromal CFUs also expressed three somatic stem cell markers: sal-like 4, CD133, and Musashi-1. Endometriotic stromal cells derived from large CFUs could differentiate into four mesenchymal lineages when cultured in the respective inducing-media, as determined by histochemical staining and RT-PCR of lineage specific markers. These findings demonstrate that ovarian endometrioma contains a subset of cells displaying somatic stem cell properties.

Putative stem cell activity of human endometrial epithelial and stromal cells during the menstrual cycle.

OBJECTIVE: To determine whether menstrual cycle stage or activity has an effect on the clonogenic activity of human endometrial epithelial and stromal cells. DESIGN: Clonal analysis of human endometrial epithelial and stromal cells derived from full-thickness endometrium. SETTING: University research laboratory. PATIENT(S): Twenty-six women of varying age and race undergoing hysterectomy for nonendometrial pathologies. INTERVENTION(S): Full-thickness human endometrial tissue was dissociated into single cells. Epithelial and stromal cells were separated using magnetic beads, and cloning assays were performed in serum-containing or growth factor-supplemented serum-free medium. MAIN OUTCOME MEASURE(S): Clonogenic activity of epithelial and stromal cells. RESULT(S): Clonogenicity of epithelial and stromal cells did not vary significantly between proliferative, secretory, and inactive endometrium. However, epithelial and stromal cells did show a trend for greater numbers of clonogenic cells in secretory and proliferative endometrium respectively. A large variation between samples was observed, which may have masked any significant differences. CONCLUSION(S): We found that clonogenicity does not vary from the proliferative to secretory stage of the menstrual cycle, or between active, cycling and inactive endometrium for both epithelial and stromal cells. We have demonstrated for the first time that inactive endometrium contains clonogenic epithelial and stromal cells.