J Immunoassay Immunochem. 2018 May 9. doi: 10.1080/15321819.2018.1447487. [Epub ahead of print]

Development of a novel explant culture method for the isolation of mesenchymal stem cells from human breast tumor.

Sineh Sepehr K¹, Razavi A¹, Saeidi M², Mossahebi-Mohammadi M³, Abdollahpour-Alitappeh M⁴, Hashemi SM^5,6.

Abstract

BACKGROUND:

Mesenchymal stem cells (MSCs) were isolated from various sources, including various types of tumors. However choosing an appropriate isolation method is an important step in obtaining cells with optimal quality and yield in companion with economical considerations. The purpose of this study was to isolate of more pure MSCs from human breast tumor tissue by a modified explant culture method.

METHODS AND MATERIALS:

The tumor tissues (n = 8) was cut into 1 to 3-mm cube-like pieces (explant). Each explant was placed in a well of 24-well format plates, cultured in Dulbecco's Modified Eagle's medium (DMEM), and maintained at 37°C with 5% humidified incubator. Morphological phenotypes of the cells were surveyed by an inverted microscope and wells with rather homogenous fibroblast-like morphology cell were considered as positive and selected for more expansion and characterization.

RESULTS:

A total of 185 wells, 63.7% of wells were positive that were chosen for expansion. Flowcytometry analysis demonstrated that isolated cells were positive for, CD73, CD44, CD29, CD105 and CD90 but negative for, CD11b, CD45, CD34, and HLA‑DR. In addition, cells possessed the capability of multipotential differentiation into osteoblasts and adipocytes.