Hormonal oral contraceptive influence on isolation, Characterization and cryopreservation of mesenchymal stem cells from menstrual fluid

Abstract

The purpose of the study was to evaluate whether the intake of hormonal oral contraceptive influences the viability of mesenchymal stem cell. Sixteen healthy female volunteers with regular menstrual cycles were invited to participate. Menstrual fluid was collected on the day of maximum flux, and collected cells were analyzed by a ‘minimal standard’ for MSC characterization: plastic adherence, trilineage (adipogenic, osteogenic, chondrogenic) in vitro differentiation and a minimalistic panel of markers assessed by flow cytometry (CD731, CD901, CD1051, CD34–, CD45–) using monoclonal antibodies. The participants were divided into two groups: Group 1 – no hormonal contraceptive use; Group 2 – hormonal oral contraceptive use. The median of the menstrual fluid volume was 5.0 and the median number of cells was 5.2 × 10⁶. Median of cell viability was 89.3%. After culture, mesenchymal stem cells increased from 0.031% of the total cells to 96.9%. The cells formed clusters and reached confluence after 15–21 days of culture in the first passage. In the second passage, clusters and the confluence were observed after 3 days of culture. No difference was observed between the groups. Our data suggest that oral hormonal contraceptive intake maintains the viability of mesenchymal stem cells from menstrual fluid.

摘要

本研究的目的是评估激素口服避孕药的摄入是否会影响间充质干细胞的活力。本研究邀请16名有规律月经周期的健康女性志愿者参加。在月经量最大的当天收集经血, 并通过用于MSC表征的“最小标准”分析收集的细胞：塑料粘附, 三线（成脂, 成骨, 软骨形成）体外分化和应用单克隆抗体进行的流式细胞术评估的简略标记组 （CD731, CD901, CD1051, CD34-, CD45-）。受试者分成两组：第1组-不用激素避孕药；第2组-应用激素口服避孕药。经血体积中位数为5.0, 细胞中位数为5.2 × 10⁶。细胞活力中位数为89.3%。培养后, 间充质干细胞从总细胞的0.031%增加到96.9%。在第一次传代培养15-21天后, 细胞形成簇达到融合。第二代培养3天后细胞形成簇融合。两组之间无差异。我们的数据表明口服激素避孕药可以维持经血间充质干细胞的活力。

The Chinese abstracts are translated by Prof. Dr. Xiangyan Ruan and her team: Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, China.

Keywords:

• Stem cell
• mesenchymal stromal cells
• menstrual fluid
• hormonal oral contraception

Acknowledgments

The authors thank the subjects for their participation and for their informed written consent.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

Cryopraxis Criobiologia Ltda. sponsored this study.