Differentiation of Human Breast-Milk Stem Cells to Neural Stem Cells and Neurons

Abstract

Objectives. Human breast milk contains a heterogeneous population of cells that have the potential to provide a noninvasive source of cells for cell therapy in many neurodegenerative diseases without any ethical concern. The objectives of this study were to differentiate the breast milk-derived stem cells (BMDSC) toward neural stem cells and then into the neurons and neuroglia. Materials and Methods. To do this, the BMDSC were isolated from human breast milk and cultured in Dulbecco's modified Eagle medium/F12 (DMEM/F12) containing fibroblast growth factor (bFGF). The cells were then characterized by evaluation of the embryonic and stem cell markers. Then, the cells were exposed to culture medium containing 1% B27 and 2% N2 for 7–10 days followed by medium supplemented with B27, N2, bFGF 10 µg/mL, and endothelial growth factor (EGF) 20 µg/mL. Then, the sphere-forming assay was performed. The spheres were then differentiated into three neural lineages by withdrawing growth factor in the presence of 5% FBS (fetal bovine serum). The immunofluorescence was done for β-tubulin III, O4, and GFAP (glial fibrillary acidic protein). Results. The results indicated that the cells expressed both embryonic and mesenchymal stem cell (MSC) markers. They also showed neurospheres formation that was nestin-positive. The cells were also differentiated into all three neural lineages. Conclusion. The BMDSC can behave in the same way with neural stem cells. They were differentiated into oligodendrocytes, and astrocytes as well as neurons.

Novel DRAQ5™/SYTOX® Blue Based Flow Cytometric Strategy to Identify and Characterize Stem Cells in Human Breast Milk

“…Most reports characterize breast milk cells using flow cytometry after several days of culturing to discriminate cells from cellular fragments and debris in human breast milk, but cell proliferation and differentiation is influenced by differing culturing methods (7)(8)(9)(10)(11). At present only few studies analyzed the cell content in freshly isolated human breast milk cells by flow cytometry (12)(13)(14): Hoechst 33342 staining was used to define a small side-population of cells (12), and detection of cell surface markers to characterize the hematopoietic, mesenchymal, myoepithelial, and immune cell content in freshly isolated breast milk (13).…”

Optimization of time for neural stem cells transplantation for brain stroke in rats

“…Three days later, the NSCs were differentiated into neurons, oligodendrocytes and astrocytes. To confirm the differentiation of the NSCs, immunocytochemistry was performed for β-tubulin III (neuron marker), glial fibrillary acidic protein (Gfap; an astrocyte marker) and Oligodendrocyte marker Olig2 as described earlier (18).…”

“…Immunostaining of cells for specific markers was essentially carried out according to our standard protocols as described earlier (18). Briefly, the cells were cultured on glass slides and fixed with 4% paraformaldehyde for 20 minutes at 4 ℃.…”

“…The cells were then washed ×3 with PBS and respective incubated with fluorescent-conjugated secondary antibodies for 45 minutes at room temperature. The nuclei were labeled with 4,6-diamino-2-phenylindole dihydrochloride (DAPI; Millipore Cat #S7113, 1:1,000) as described earlier (18). The samples were later fixed and visualized under fluorescence microscope (Olympus BX53 Japan) fitted with camera and software Cell-sens.…”

Mleko kobiece jako naturalny produkt leczniczy

“…Oba typy komórek różnicują w zakresie trzech listków zarodkowych. Oznacza to, że sutek jest gruczołem wydzielającym laktocyty, komórki mioepitelialne oraz jeszcze niezróżnicowane pluripotencjalne komórki macierzyste, które mogą zostać przekształcone w komórki neuronalne, glejowe, osteoblasty, chondrocyty, adipocyty i kardiomiocyty, a także hepatocyty i komórki trzustki [18]. Ponadto, podawanie hBSCs myszom nie spowodowało tworzenia potworniaków, co czyni je wyjątkowo atrakcyjnymi w aspekcie praktycznego wykorzystania w terapii [19].…”