Posted on July 21, 2021
Hassiotou and co-workers recently demonstrated the isolation of cell populations from individual breasts dairy, which expressed stem cell-related transcription factors such as Oct4, Sox2 and Nanog and proved their multilineage differentiation potential 
Hassiotou and co-workers recently demonstrated the isolation of cell populations from individual breasts dairy, which expressed stem cell-related transcription factors such as Oct4, Sox2 and Nanog and proved their multilineage differentiation potential . A less popular marker for the characterization of mammary stem cells is nestin, which has been expressed in cells of the mammary gland [23,24]. the isotype control (green line) for each measurement. PE, phycoerythrin; PerCP, peridinin chlorophyll protein. scrt229-S2.tiff (414K) GUID:?494453A8-8469-4F5E-9CCB-0C854E4E3D49 Abstract Introduction Nestin-expressing cells isolated from different human tissues reveal self-renewal capacity and a multilineage differentiation potential. In particular, adult stem/progenitor cell populations from exocrine glands such as the pancreas, salivary gland and sweat gland are characterized by prominent nestin expression. Interestingly, human mammary gland histological examinations also demonstrated the existence of PR-619 nestin-positive cells in the ductal compartments. Within the scope of our previous work we wonder whether an isolation of nestin-positive cell populations from human mammary gland biopsies is Rabbit polyclonal to KLK7 possible and what characteristics they have culture, the obtained cell populations are characterized by their prominent nestin expression. The cells share surface proteins commonly expressed on adult stem cells. We demonstrated the expression of stem cell-related genes like Oct4, Sox2, KLF4 and Nanog, and confirmed multipotent differentiation capacity by detecting transcripts expressed in endodermal, mesodermal and ectodermal cell types. Conclusion With this study we present an efficient procedure for isolation and propagation of nestin-positive stem cells obtained from male and female breast tissue, which is frequently available. The established multipotent cell populations could be easily expanded and thus hold promise for cell-based therapies and personalized medicine. and assays of mammary-derived cell populations had been performed. First insights into the characteristics and behavior of isolated mammary stem cells were gained from adherent two-dimensional cultures or from suspension cultures; for example, as mammospheres. The focus of gene and protein expression analysis for those studies was chosen for surface markers and/or cytokeratin synthesis, to classify the state of differentiation in comparison with histological data [11,13-21]. The results demonstrated that the mammary stem cells preserve their capability for differentiation into mammary cells and retain the sensitivity against tissue-specific hormones [16,22]. On the contrary, their continuous proliferation and the expression of stem cell and proliferation-related markers such as Ki67, musashi-1, Sca-1 or p21 affirmed that they keep a stem/progenitor state [11,13-18]. Interestingly, even the isolation and cultivation of stem cells from human breast milk has been documented [19-21]. Hassiotou and colleagues recently demonstrated the isolation of cell populations from human breast milk, which expressed stem PR-619 cell-related transcription factors such as Oct4, Sox2 and Nanog and proved their multilineage differentiation potential . A less popular marker for the characterization of mammary stem cells is nestin, which has been expressed in cells of the mammary gland [23,24]. The expression of the intermediate filament nestin has already been shown to be associated with multipotency and stemness of several cell populations . We and others demonstrated that different glandular tissues (for example, pancreas, salivary glands or sweat glands) yield nestin-positive stem cell populations with multipotent, long-term proliferative potential and the pellet was resuspended in DMEM (Invitrogen, Darmstadt, Germany) with 20% (v/v) FCS (PAA, C?lbe, Germany) and penicillin/streptomycin (PAA, C?lbe, Germany). The acini-containing medium was seeded into one well of a six-well cell culture test plate (TPP, Trasadingen, Switzerland) and this primary culture was incubated for 2 days in a humidified incubator with 37C and 5% CO2 in the atmosphere. After 2 days the first media exchange was performed and the cultivated cells were propagated until highly confluent colonies were visible. Cultivation of mammary cells The cultivation after the first trypsinization was performed with DMEM with 10% (v/v) FCS and penicillin/streptomycin in TPP cell culture plastic. We generally used polystyrene-plastic dishes as delivered (all cell culture treated by the supplier TPP). The incubator settings were the same as already described. When the population reached a confluence of >80% covered growth area, it was split in a ratio of 1 1:3 regarding the growth area by standard trypsinization. This includes the removal of the old medium, one washing step with PBS (Invitrogen, Darmstadt, Germany) and incubation for 2 minutes at 37C after adding the TrypsinCethylenediamine tetraacetic acid mixture (PAA, PR-619 C?lbe, Germany). By microscopic control of the cells detachment, the stopping procedure starts by adding the doubled volume media to the trypsin-based suspension. The whole liquid was afterwards centrifuged at 180??for 5 minutes and the pellet was resuspended with cultivation medium. The cells were then reseeded.