A Novel Xeno-Free Method to Isolate Human Endometrial Mesenchymal Stromal Cells (E-MSCs) in Good Manufacturing Practice (GMP) Conditions
| Autor: | Stefano Canosa, Katia Mareschi, Elena Marini, Andrea Roberto Carosso, Sara Castiglia, Deborah Rustichelli, Ivana Ferrero, Gianluca Gennarelli, Benedetta Bussolati, Alberto Nocifora, Valentina Asnaghi, Massimiliano Bergallo, Ciro Isidoro, Chiara Benedetto, Alberto Revelli, Franca Fagioli |
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| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
Adult
Cell Culture Techniques Bone Marrow Cells Endometrial mesenchymal stromal cells Asherman’s syndrome Endometrial sampling Endometrial thickness Good Manufacturing Practice (GMP) Human platelet lysate (HPL) Infertility Catalysis Inorganic Chemistry Endometrium Young Adult Humans Physical and Theoretical Chemistry Molecular Biology Cells Cultured Spectroscopy Cell Proliferation Organic Chemistry Cell Differentiation Mesenchymal Stem Cells General Medicine Computer Science Applications endometrial mesenchymal stromal cells infertility endometrial thickness human platelet lysate (HPL) endometrial sampling Female Biomarkers |
| Zdroj: | International Journal of Molecular Sciences; Volume 23; Issue 4; Pages: 1931 |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms23041931 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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