A Novel Xeno-Free Method to Isolate Human Endometrial Mesenchymal Stromal Cells (E-MSCs) in Good Manufacturing Practice (GMP) Conditions

• Published in: International journal of molecular sciences Vol. 23; no. 4; p. 1931
• Main Authors: Canosa, Stefano, Mareschi, Katia, Marini, Elena, Carosso, Andrea Roberto, Castiglia, Sara, Rustichelli, Deborah, Ferrero, Ivana, Gennarelli, Gianluca, Bussolati, Benedetta, Nocifora, Alberto, Asnaghi, Valentina, Bergallo, Massimiliano, Isidoro, Ciro, Benedetto, Chiara, Revelli, Alberto, Fagioli, Franca
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 09.02.2022; MDPI
• Subjects: Adipocytes; Adult; Asherman’s syndrome; Biomarkers - metabolism; Biopsy; Bone marrow; Bone Marrow Cells - cytology; Cell culture; Cell Culture Techniques - methods; Cell Differentiation - physiology; Cell Proliferation - physiology; Cells, Cultured; Curettage; Differentiation; endometrial mesenchymal stromal cells; endometrial thickness; Endometrium; Endometrium - cytology; Endometrium - metabolism; Female; Flow cytometry; Gene expression; Good Manufacturing Practice; Good Manufacturing Practice (GMP); Gynecological diseases; human platelet lysate (HPL); Humans; In vitro fertilization; Infertility; Invasiveness; Karyotypes; Manufacturing; Menstruation; Mesenchymal Stem Cells - cytology; Mesenchyme; Morphology; Phenotypes; Sampling; Sampling methods; Stem cells; Stromal cells; Young Adult; human platelet lysate (HPL); endometrial mesenchymal stromal cells; endometrial thickness; Asherman’s syndrome; Good Manufacturing Practice (GMP); endometrial sampling; infertility
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23041931

The cyclic regeneration of human endometrium is guaranteed by the proliferative capacity of endometrial mesenchymal stromal cells (E-MSCs). Due to this, the autologous infusion of E-MSCs has been proposed to support endometrial growth in a wide range of gynecological diseases. We aimed to compare two different endometrial sampling methods, surgical curettage and vacuum aspiration biopsy random assay (VABRA), and to validate a novel xeno-free method to culture human E-MSCs. Six E-MSCs cell samples were isolated after mechanical tissue homogenization and cultured using human platelet lysate. E-MSCs were characterized for the colony formation capacity, proliferative potential, and multilineage differentiation. The expression of mesenchymal and stemness markers were tested by FACS analysis and real-time PCR, respectively. Chromosomal alterations were evaluated by karyotype analysis, whereas tumorigenic capacity and invasiveness were tested by soft agar assay. Both endometrial sampling techniques allowed efficient isolation and expansion of E-MSCs using a xeno-free method, preserving their mesenchymal and stemness phenotype, proliferative potential, and limited multi-lineage differentiation ability during the culture. No chromosomal alterations and invasive/tumorigenic capacity were observed. Herein, we report the first evidence of efficient E-MSCs isolation and culture in Good Manufacturing Practice compliance conditions, suggesting VABRA endometrial sampling as alternative to surgical curettage.