Rapid selection of mesenchymal stem and progenitor cells in primary prostate stromal cultures

Abstract The hypothalamus is a key homeostatic brain region and the primary effector of neuroendocrine signaling. Recent studies show that early embryonic developmental disruption of this region can lead to neuroendocrine conditions later in life, suggesting that hypothalamic progenitors might be sensitive to exogenous challenges. To study the behavior of hypothalamic neural progenitors, we developed a novel dissection methodology to isolate murine hypothalamic neural stem and progenitor cells at the early timepoint of embryonic day 12.5, which coincides with peak hypothalamic neurogenesis. Additionally, we established and optimized a culturing protocol to maintain multipotent hypothalamic neurospheres that are capable of sustained proliferation or differentiation into neurons, oligodendrocytes, and astrocytes. We characterized media requirements, appropriate cell seeding density, and the role of growth factors and sonic hedgehog (Shh) supplementation. Finally, we validated the use of fluorescence activated cell sorting of either Sox2GFPKI or Nkx2.1GFPKI transgenic mice as an alternate cellular isolation approach to enable enriched selection of hypothalamic progenitors for growth into neurospheres. Combined, we present a new technique that yields reliable culturing of hypothalamic neural stem and progenitor cells that can be used to study hypothalamic development in a controlled environment.

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Fluorescence-Based Selection of Gene-Corrected Hematopoietic Stem and Progenitor Cells Based on Acid Sphingomyelinase Expression

Methods in Enzymology - Sphingolipid Metabolism and Cell Signaling, Part B ◽

10.1016/s0076-6879(00)12919-2 ◽

2000 ◽

pp. 330-338

Author(s):

Edward H. Schuchman ◽

Shai Erlich ◽

Silvia R.P. Miranda ◽

Tama Dinur ◽

Arie Dagan ◽

...

Keyword(s):

Progenitor Cells ◽

Acid Sphingomyelinase ◽

Hematopoietic Stem ◽

Stem And Progenitor Cells ◽

Selection Of

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Oestrogen action on thyroid progenitor cells: relevant for the pathogenesis of thyroid nodules?

Journal of Endocrinology ◽

10.1530/joe-13-0029 ◽

2013 ◽

Vol 218 (1) ◽

pp. 125-133 ◽

Cited By ~ 27

Author(s):

Shuhang Xu ◽

Guofang Chen ◽

Wen Peng ◽

Kostja Renko ◽

Michael Derwahl

Keyword(s):

Progenitor Cells ◽

Thyroid Nodules ◽

Cellular Heterogeneity ◽

Sodium Iodide Symporter ◽

Thyroid Cells ◽

Proliferative Effect ◽

Supportive Role ◽

Stem And Progenitor Cells ◽

Selection Of

Benign and malignant thyroid nodules are more prevalent in females than in males. Experimental data suggest that the proliferative effect of oestrogen rather than polymorphisms is responsible for this gender difference. This study analysed whether both differentiated thyroid cells and thyroid stem and progenitor cells are targets of oestrogen action. In thyroid stem/progenitor cells derived from nodular goitres, the ability of 17β-oestradiol (E2) to induce the formation of thyrospheres and the expression of oestrogen receptors (ERs) and the effect of E2 on the growth and expression of markers of stem cells and thyroid differentiation (TSH receptor, thyroperoxidase, thyroglobulin and sodium iodide symporter (NIS)) were analysed. E2 induced thyrosphere formation, albeit to a lower extent than other growth factors. Thyroid stem and progenitor cells expressed ERα (ESR1) and ERβ (ESR2) with eight times higher expression levels of ERα mRNA compared with the differentiated thyrocytes. E2 was a potent stimulator of the growth of thyroid stem/progenitor cells. In contrast, TSH-induced differentiation of progenitor cells, in particular, the expression of NIS, was significantly inhibited by E2. In conclusion, oestrogen stimulated the growth and simultaneously inhibited the differentiation of thyroid nodule-derived stem/progenitor cells. From these data and based on the concept of cellular heterogeneity, we hypothesize a supportive role of oestrogen in the propagation of thyroid stem/progenitor cells leading to the selection of a progeny of growth-prone cells with a decreased differentiation. These cells may be the origin of hypofunctioning or non-functioning thyroid nodules in females.

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Integrated Colony Imaging, Analysis, and Selection Device for Regenerative Medicine

SLAS TECHNOLOGY Translating Life Sciences Innovation ◽

10.1177/2211068216676587 ◽

2016 ◽

Vol 22 (2) ◽

pp. 217-223 ◽

Cited By ~ 3

Author(s):

Edward Kwee ◽

Edward E. Herderick ◽

Thomas Adams ◽

James Dunn ◽

Robert Germanowski ◽

...

Keyword(s):

Stem Cells ◽

Progenitor Cells ◽

Cell Types ◽

Automated Image Analysis ◽

Adherent Cell ◽

Imaging Analysis ◽

Cell Therapies ◽

Stem And Progenitor Cells ◽

Cell Sources ◽

Selection Of

Stem and progenitor cells derived from human tissues are being developed as cell sources for cell-based assays and therapies. However, tissue-derived stem and progenitor cells are heterogeneous. Differences in observed clones of stem cells likely reflect important aspects of the underlying state of the source cells, as well as future potency for cell therapies. This paper describes a colony analysis and picking device that provides quantitative analysis of heterogeneous cell populations and precise tools for cell picking for research or biomanufacturing applications. We describe an integrated robotic system that enables image acquisition and automated image analysis to be coupled with rapid automated selection of individual colonies in adherent cell cultures. Other automated systems have demonstrated feasibility with picking from semisolid media or off feeder layers. We demonstrate the capability to pick adherent bone-derived stem cells from tissue culture plastic. Cells are efficiently picked from a target site and transferred to a recipient well plate. Cells demonstrate viability and adherence and maintain biologic potential for surface markers CD73 and CD90 based on phase contrast and fluorescence imaging 6 days after transfer. Methods developed here can be applied to the study of other stem cell types and automated culture of cells.

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