Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Validation of a Novel Xeno-Free Method for Human Endometrial Mesenchymal Stromal Cells (E-MSCs) Isolation and Culture

Stefano Canosa
ORCID logo ,
Katia Mareschi
* ORCID logo ,
Elena Marini
,
Andrea Roberto Carosso
ORCID logo ,
Sara Castiglia
,
Deborah Rustichelli
,
Ivana Ferrero
,
Gianluca Gennarelli
,
Benedetta Bussolati
ORCID logo ,
Alberto Nocifora
,
Valentina Asnaghi
,
Massimiliano Bergallo
,
Ciro Isidoro
ORCID logo ,
Chiara Benedetto
,
Alberto Revelli
,
Franca Fagioli
Version 1 : Received: 11 March 2021 / Approved: 12 March 2021 / Online: 12 March 2021 (20:51:55 CET)

How to cite: Canosa, S.; Mareschi, K.; Marini, E.; Carosso, A.R.; Castiglia, S.; Rustichelli, D.; Ferrero, I.; Gennarelli, G.; Bussolati, B.; Nocifora, A.; Asnaghi, V.; Bergallo, M.; Isidoro, C.; Benedetto, C.; Revelli, A.; Fagioli, F. Validation of a Novel Xeno-Free Method for Human Endometrial Mesenchymal Stromal Cells (E-MSCs) Isolation and Culture. Preprints 2021, 2021030354. https://doi.org/10.20944/preprints202103.0354.v1 Canosa, S.; Mareschi, K.; Marini, E.; Carosso, A.R.; Castiglia, S.; Rustichelli, D.; Ferrero, I.; Gennarelli, G.; Bussolati, B.; Nocifora, A.; Asnaghi, V.; Bergallo, M.; Isidoro, C.; Benedetto, C.; Revelli, A.; Fagioli, F. Validation of a Novel Xeno-Free Method for Human Endometrial Mesenchymal Stromal Cells (E-MSCs) Isolation and Culture. Preprints 2021, 2021030354. https://doi.org/10.20944/preprints202103.0354.v1

Abstract

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, the surgical curettage and the Vacuum Aspiration Biopsy Random Assay, and to validate a novel xeno-free method to culture human E-MSCs. Six E-MSCs cell lines were isolated after a 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 was 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 to efficiently isolate and expand E-MSCs using a xeno-free method preserving their mesenchymal and stemness phenotype, proliferative potential and 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.

Keywords

Endometrial Mesenchymal Stromal Cells; Good Manufacturing Practice (GMP); infertility; Asherman’s syndrome, endometrial thickness; Human platelet lysate (HPL); endometrial sampling

Subject

Biology and Life Sciences, Anatomy and Physiology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Download PDF

Comments (0)

We encourage comments and feedback from a broad range of readers. See criteria for comments and our Diversity statement.

Leave a public comment
Send a private comment to the author(s)
* All users must log in before leaving a comment
Views 0
Downloads 0
Comments 0
Metrics 0
Get PDF Cite Share 0
Bookmark BibSonomy Mendeley Reddit Delicious
×
Alerts
Notify me about updates to this article or when a peer-reviewed version is published.
We use cookies on our website to ensure you get the best experience.
Read more about our cookies here.