Transcriptomic profiling analysis of human endometrial stromal cells treated with autologous platelet-rich plasma

Abstract

Purpose: To clarify the mechanisms of intrauterine platelet-rich plasma (PRP) infusion that support embryo implantation in in vitro fertilization treatment.

Methods: Blood and endometrial samples were collected from four infertile women. Human endometrial stromal cells (HESCs) were cultured and passaged equally into four cell culture dishes in each patient. Two were treated with PRP twice, and the other two were treated with vehicle. Subsequently, two cultures with and without PRP were decidualized with 8-bromoadenosine 3',5'-cyclic AMP and progesterone for 5 days.

Results: The gene expression in undifferentiated or decidualized HESCs with and without PRP was compared. In the microarray analysis, 381 and 63 differentially expressed genes were detected in undifferentiated and decidualized HESCs, respectively. In the undifferentiated HESCs, PRP was found to promote the gene expression associated with cell growth, tissue regeneration, proinflammatory response, and antibiotic effects. In decidualized HESCs, PRP was found to attenuate the gene expression involved in cell proliferation and inflammation by inhibiting the expression of phosphoinositide 3-kinase signaling.

Conclusions: Platelet-rich plasma regulates the reprogramming of cell proliferation and inflammation depending on menstrual cycle phases in an appropriate manner, suggesting that PRP has the potential to increase endometrial thickness in the proliferative phase and improve immune tolerance in the secretory phase.

Keywords: cell growth; decidualization; endometrium; phosphoinositide 3‐kinase signaling pathway; platelet‐rich plasma.

FIGURE 1

Cell proliferation in undifferentiated human endometrial stromal cells (HESCs) with and without platelet‐rich plasma (PRP) treatment. (A) Cell proliferation in the strips after endometrial scratching in undifferentiated HESCs treated with and without PRP. (B) Number and increasing rate in the strips in eight independent primary cultures from four patients at 1 and 3 days after endometrial scratching. The data are expressed as mean fold change ± SEM. The dates are counted from the day of endometrial scratching.

FIGURE 2

Platelet‐rich plasma (PRP)‐affected genes in undifferentiated and decidualized human endometrial stromal cells (HESCs). (A) Venn diagram showing 381 and 63 PRP‐affected genes were extracted in undifferentiated and decidualized HESCs, respectively. Ten genes were identified in both undifferentiated and decidualized HESCs. (B) Principal component analysis in undifferentiated and decidualized HESCs with and without PRP. Blue and green plots show the samples with and without PRP treatment, respectively. Triangle and square plots show the samples of undifferentiated and decidualized HESCs, respectively. (C) Heatmap analysis in undifferentiated HESCs with and without PRP (D) Heatmap analysis in decidualized HESCs with and without PRP.

FIGURE 3

Platelet‐rich plasma (PRP)‐affected genes in the PI3K/AKT signaling pathway in decidualized human endometrial stromal cells (HESCs). In decidualized HESCs treated with and without platelet‐rich plasma, inclusive real‐time quantitative polymerase chain reaction (RTQ‐PCR) array analysis of the PI3K/AKT signaling pathway demonstrated 10 significantly upregulated genes and 10 significantly downregulated genes, respectively, using a cutoff value of p < 0.05.

FIGURE 4

Therapeutic effects of intrauterine platelet‐rich plasma (PRP) infusion in proliferative and secretory phases. Intrauterine PRP infusion promotes tissue repair with transient inflammatory response, cell growth and migration, and antibiotic effect in undifferentiated human endometrial stromal cells, whereas cell proliferation and the inflammatory immune response are attenuated during decidualization of the endometrium. Depending on the menstrual cycle phases, PRP can regulate the reprogramming of inflammation and cell proliferation in an appropriate manner to support embryo implantation.