ORIGINAL ARTICLES   

Minerva Biotecnologica 2016 December;28(4):208-18

Copyright © 2016 EDIZIONI MINERVA MEDICA

lingua: Inglese

Purification and identification of CD146 positive endometrial somatic stem cell from human normal endometrium biopsy and gene expression criteria

Saeed HEIDARI-KESHEL ¹, Mostafa REZAEI-TAVIRANI ¹, Jafar AI ², Masoud SOLEIMANI ³, Maryam EBRAHIMI ¹, Alireza BARADARAN-RAFII ⁴, Saeid RAHMAN ZADEH ¹, Roghiyeh OMIDI ¹, Reza ROOZAFZOON ¹, Hakimeh ZALI ¹, Zinat GHANBARI ⁵, Ahad KHOSHZABAN ⁶

¹ Proteomics Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran; ² Tissue Engineering Department, Faculty of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran; ³ Hematology Department, Tarbbiat Modares University, Tehran, Iran; ⁴ Ocular Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran; ⁵ Enzyme Technology Lab, Genetics and Metabolism Research Group, Pasteur Institute of Iran, Tehran, Iran; ⁶ Iranian Tissue Bank Research Center, Tehran University of Medical Sciences, Tehran, Iran

BACKGROUND: The endometrial is answerable for the sex steroid hormones, endures tremendous growth in a cyclic manner, and is renewed and shed during a woman’s lifetime. It has been recommended that the human endometrium may contain a population of stem cells that is responsible for its striking ability to regenerate. The aim of this study was the isolation and characterization of sub population stem cell in vitro condition from healthy donors.
METHODS: Purification of endometrial stromal stem cell was acquired by selecting cells with magnetic bead sorting. The surface markers of stem cell were discerned by flow cytometry. Proliferation of endometrial stem cell was ascertained by MTT assay. The osteogenic potential was assessed by alizarin red staining, the adipogenic potential by oil red O staining and karyotyping were accomplished on human endometrial somatic stem cell. Then, appraised the specific genes of the osteogenic, adipogenic and stem cell were assessed by reverse transcription polymerase chain reaction (RT-PCR) and real time PCR method.
RESULTS: Our data displayed that these cells had a negative expression of SSEA1, STAT3, CD34, CD45, CD133 and BMP4. On the other hand, they had a positive expression for Pou5f1, Nanog, Rex1, CD105, CD146, CD90, CD73 and B2m. Moreover, CD146 positive endometrial somatic stem cell showing normal karyotype is their ability to be continuously cultured for more than 20 passages. This cell has been differentiated to osteogenic and Adipogenic lineage.
CONCLUSIONS: This study illustrates the presence of CD146 positive human endometrial somatic stem cell in the human endometrium and reveals their potential use in regenerative medicine and taming stem cell heterogeneity for cell therapy and tissue engineering, like in the treatment of related infertility diseases.