Safety of Intraovarian Injection of Human Mesenchymal Stem Cells in a Premature Ovarian Insufficiency Mouse Model

Abstract

Primary ovarian insufficiency (POI), a condition in which there is a loss of ovarian function before the age of 40 years, leads to amenorrhea and infertility. In our previously published studies, we demonstrated recovery of POI, correction of serum sex hormone levels, increase in the granulosa cell population, and restoration of fertility in a chemotherapy-induced POI mouse model after intraovarian transplantation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs). While hBM-MSC may be a promising cell source for treatment of POI, there are few reports on the safety of stem cell-based therapy for POI. For future clinical applications, the safety of allogenic hBM-MSCs for the treatment of POI through intraovarian engraftment needs to be addressed and verified in appropriate preclinical models. In this study, we induced POI in C57/BL6 mice using chemotherapy, then treated the mice with hBM-MSCs (500,000 cells/ovary) by intraovarian injection. After hBM-MSC treatment, we analyzed the migration of engrafted cells by genomic DNA polymerase chain reaction (PCR) using a human-specific ALU repeat and by whole-body sectioning on a cryo-imaging system. We examined the possibility of transfer of human DNA from the hBM-MSCs to the resulting offspring, and compared the growth rate of offspring to that of normal mice and hBM-MSC-treated mice. We found that engrafted hBM-MSCs were detected only in mouse ovaries and did not migrate into any other major organs including the heart, lungs, and liver. Further, we found that no human DNA was transferred into the fetus. Interestingly, the engrafted cells gradually decreased in number and had mostly disappeared after 4 weeks. Our study demonstrates that intraovarian transplantation of hBM-MSCs could be a safe stem cell-based therapy to restore fertility in POI patients.

Mesenchymal stem cell therapy ameliorates metabolic dysfunction and restores fertility in a PCOS mouse model through interleukin-10

“…In our previous study, we showed the presence of injected human MSC cells in to the ovarian tissue by immune fluorescence assay [ 67 ]. Our recent study also suggested that engrafted MSC by intra-ovarian injection stayed in to the ovary for more than 2 weeks [ 68 ]. Our PCR-based quantification analysis for engrafted human MSC shows that approximately 30% of cells are still present in the ovary for 2 weeks after injection and later gradually degraded [ 68 ].…”

“…Our recent study also suggested that engrafted MSC by intra-ovarian injection stayed in to the ovary for more than 2 weeks [ 68 ]. Our PCR-based quantification analysis for engrafted human MSC shows that approximately 30% of cells are still present in the ovary for 2 weeks after injection and later gradually degraded [ 68 ]. Due to limited lifespan of the engrafted MSC, it is not possible to expect a long-term treatment by only single injection of MSC.…”

“…Second is lack of direct evidence of MSC engraftment and inflammatory infiltration in the ovary tissue. In our previous study, we already reported that engrafted hMSCs are found in the ovary [ 67 ] and stayed in ovary more than 2 weeks [ 68 ]. Other previous studies also reported that decreased population of immune cells [ 70 ].…”

Research Progress on the Treatment of Premature Ovarian Failure Using Mesenchymal Stem Cells: A Literature Review

“…, Lai et al, Song et al and Park et al . also demonstrated that after transplantation, MSCs home to damaged tissue and reach the site of injured ovaries ( Liu et al, 2014 ; Lai et al, 2015 ; Jalalie et al, 2019 ; Park et al, 2021 ). However, studies have also shown that although MSCs have a homing effect, they cannot directly differentiate into oocytes but do localize in the ovarian matrix, secrete various cytokines and improve ovarian reserve function through the paracrine pathway ( Takehara et al, 2013 ; Gabr et al, 2016 ; Li et al, 2017 ).…”

Extracellular Vesicles Released from Neprilysin Gene-Modified Human Umbilical Cord-Derived Mesenchymal Stem Cell Enhance Therapeutic Effects in an Alzheimer’s Disease Animal Model

“…We produced the mixed populations by diluting hUC-MSCs to the cells of the normal mouse (hUC-MSCs compared to normal mouse cells: 100%, 50%, 5%, 0.5%, and 0%, and the total number of cells remains constant at 5 × 10 5 ). We performed qRT-PCR for human-specific Alu sequences with 25 ng of DNA extracted in dilution groups and generated an Alu standard curve (calculation curve) and equation [48]. Subsequently, we estimated the number of human cells contained within mouse brain in each group by substituting the Ct value of the experimental group sample in the standard curve and equation.…”