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Neural Differentiation of Rodent Neural Progenitor Cells and Isolation and Enrichment of Human Neural Progenitor/Stem Cells

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Abstract

Neural progenitor cells (NPC) are multipotent and give rise to neurons, astrocytes and oligodendrocytes. NPC possess the ability of in vivo and in vitro proliferation and can therefore be expanded for research proposes. Cell proliferation generates a three dimensional aggregate called neurospheres; posteriorly, these neurospheres can be differentiated into multiple cell types and become an important tool for understanding the mechanisms regarding cell differentiation and neuronal regeneration. In this chapter, we describe the extraction of rat neural progenitor cells from embryonic telencephalon (14 days) and generation of neurospheres. Next, we describe protocols to isolate and proliferate human fetal stem cells derived from elective abortion, recognizing the ethical dilemmas. The advantage of using the neurospheres-rat model is that the differentiation can be achieved by removing the growth factors and allowing cell differentiation in poly-d-lysine/laminin coated plates. The cells can then be dissociated and grafted in animals or be differentiated and grafted, depending on the research hypothesis. The endogenous cells can be followed by staining with lipophilic dyes or analyzed by adding 5-Bromo-2′-deoxyuridine (BrdU) using the posterior double immunofluorescence technique, which identifies the post-mitotic neural progenitor cells and the fate acquired. The methods here described will help the researcher to perform cell extraction, differentiation without cell enrichment, track the proliferating neural progenitor cells and perform characterization after the cell graft, as well as isolate and maintain human fetal stem cells.

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Acknowledgments

This work was supported by NIH (U54NS083924) for AHM and MNG. JLRL received support from the NIGMS (8P20GM103475-12) and MBRS-RISE at UNE (2R25GM066250-05A1) from NIH.

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Authors and Affiliations

  1. Pharmacology and Toxicology Department, School of Medicine University of Puerto Rico Medical Science Campus, San Juan, PR, USA

    Antonio H. Martins

  2. Department of Biochemistry, Universidad Central del Caribe, Bayamón, PR, USA

    Antonio H. Martins & Maxine Nicole Gonzalez

  3. School of Science and Technology, Universidad Del Este, AGMUS, Carolina, PR, USA

    Jose L. Roig-Lopez

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  1. Antonio H. Martins
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  2. Jose L. Roig-Lopez
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Correspondence to Antonio H. Martins .

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Martins, A.H., Roig-Lopez, J.L., Gonzalez, M.N. (2016). Neural Differentiation of Rodent Neural Progenitor Cells and Isolation and Enrichment of Human Neural Progenitor/Stem Cells. In: Working with Stem Cells. Springer, Cham. https://doi.org/10.1007/978-3-319-30582-0_4

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