Production of functional dendritic cells from menstrual blood—a new dendritic cell source for immune therapy

Abstract

Dendritic cells (DCs) are the most professional antigen-presenting cells of the mammalian immune system. They are able to phagocytize, process antigen materials, and then present them to the surface of other cells including T lymphocytes in the immune system. These capabilities make DC therapy become a novel and promising immune-therapeutic approach for cancer treatment as well as for cancer vaccination. Many trials of DC therapy to treat cancers have been performed and have shown their application value. They involve harvesting monocytes or hematopoietic stem cells from a patient and processing them in the laboratory to produce DCs and then reintroduced into a patient in order to activate the immune system. DCs were successfully produced from peripheral, umbilical cord blood-derived monocytes or hematopoietic stem cells. In this research, we produced DCs from human menstrual blood-derived monocytes. Briefly, monocytes were isolated by FACS based on FSC vs. SSC plot from lysed menstrual blood. Obtained monocytes were induced into DCs by a two-step protocol. In the first step, monocytes were incubated in RPMI medium supplemented with 2% FBS, GM-CSF, and IL-4, followed by incubation in RPMI medium supplemented with α-TNF in the second step. Our data showed that induced monocytes had typical morphology of DCs, expressed HLA-DR, HLA-ABC, CD80 and CD86 markers, exhibited uptake of dextran-FITC, stimulated allogenic T cell proliferation, and released IL-12. These results demonstrated that menstrual blood can not only be a source of stromal stem cell but also DCs, which are a potential candidate for immune therapy.

Menstrual blood-derived mesenchymal stem cells differentiate into functional hepatocyte-like cells

“…Endometrial tissue is a prolific source of stem cells that reoccur monthly . Many studies have demonstrated that endometrial stem cells can be harvested from menstrual blood by a non-invasive method Borlongan et al, 2010;Phuc et al, 2011;Allickson and Xiang, 2012). Consistent with previous reports, the MenSCs used in our study expressed classic MSC markers (CD29, CD73, CD90, and CD105) but did not express hematopoietic stem cell markers (CD34 and CD45) and immune activation markers (HLA-DR) Borlongan et al, 2010;Phuc et al, 2011;Allickson and Xiang, 2012).…”

“…Many studies have demonstrated that endometrial stem cells can be harvested from menstrual blood by a non-invasive method Borlongan et al, 2010;Phuc et al, 2011;Allickson and Xiang, 2012). Consistent with previous reports, the MenSCs used in our study expressed classic MSC markers (CD29, CD73, CD90, and CD105) but did not express hematopoietic stem cell markers (CD34 and CD45) and immune activation markers (HLA-DR) Borlongan et al, 2010;Phuc et al, 2011;Allickson and Xiang, 2012). As mentioned by , the MenSCs are also positive for both Oct-4 and SSEA-4 markers expressed by human embryonic stem cells, which may explain their rapid expansion and multi-potent capability in this study .…”

A simple in vitro method for evaluating dendritic cell-based vaccinations

“…The phagocytic capacity was analyzed as previously described 21. Briefly, iDCs and mDCs were incubated with dextran conjugated with FITC (1 mg/mL; Sigma-Aldrich Co) in culture medium for 1 hour at 37°C, or at 4°C for the negative control.…”

“…T lymphocyte proliferation stimulated by DCs was evaluated as previously described 21. There were five experimental groups with different ratios of DCs:lymphocytes (0.25:100, 0.5:100, 1:100, 2:100, and 8:100) and three control groups with DCs + phytohemagglutinin (PHA), PHA alone, or PHA + lymphocytes.…”

“…Measurement of IL-12 was performed per a previously published study 21. Briefly, mDCs were incubated in the fresh culture medium in a 24-well plate for 24 hours.…”