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Neural Lineage Markers

By: Anne Keller-Novak, Product Manager, Sigma® Life Science, Biofiles, Vol. 8, No. 15

The ever evolving field of stem cell research has been a highly fascinating yet contested area of study. Stem cell research holds promise for many areas such as developmental biology, therapeutic and regenerative medicine, and drug discovery. Potton et al. described stem cells as undifferentiated cells capable of a) proliferation, b) selfmaintenance, c) differentiation, and d) multipotency.1 Stem cells provide a blank slate for researchers to better understand what internal and external factors drive these cells to differentiate and become committed to certain lineages or cell types.

Initially, scientists worked with two kinds of stem cells, embryonic stem cells (ESC) and adult or somatic stem cells. While ESC are pluripotent and stable cell lines have been generated for research use, controversy about their origin reduces their availability and desirability. Adult stem cells are not pluripotent, and hurdles remain in isolating and culturing them, although they may present advantages in applications involving transplantation. In 2006, Yamanaka opened another door in stem cell research by illustrating the concept of "reprogramming" cells through the generation of induced pluripotent stem cells (iPSC) using transcription factors in mouse fibroblasts.2 More recently, the process has been augmented using small molecules to coax cells into certain lineages. For example, Ding et al. were able to develop a rapid and consistent method for converting human ESC to precursor neural stem cells (NSC) through the use of small molecules SB 431542 and CHIR99021.3 In another example, Mak et al. used Dorsomorphin and SB 431542 to generate neural precursors from patient specific iPSC lines.4 With additional manipulation using Sonic Hedgehog, they were able to successfully continue differentiation into dopominergic neurons.

With this research, there comes a need for lineage markers to identify and sort certain populations of cells. Over time certain markers, or combinations thereof, have become established.5 However, as cellular subpopulations become identified and refined, new markers are always being identified. Sheikh et al. identified querkopf (QKF, aka MYST4, MORF, KAT6B) as a "stemness" marker in neural stem cells.6 Querkopf, belonging to a subclass of histone acetyltransferases, has been shown to play an essential role in normal brain development, where over-expression leads to malignancy and loss of function leads to defects.7 In the study, cells that expressed high levels of QKF generated all three neuronal cell types; astrocytes, oligodendrocytes, and neurons. Furthermore, varying levels of QKF expression allowed isolation of specific cell populations, e.g. NSC, neuroblasts, ependymal cells, and transit amplifying cells.

Cusulin et al. utilized a multitude of neuronal markers, such as Nestin, MAP2, and CD11b, in their quest to better understand the role cell fusion may play upon transplantation of NSC(s) between mature neurons or microglial cells.8 In addition, to the aforementioned markers, Sigma® Life Science offers a wide array of neural lineage markers. Prestige Antibodies® powered by Atlas Antibodies are validated in neuro tissues and cell lines. IHC images from human cerebellum, hippocampus, lateral ventricle, and cerebral cortex tissues are available for each antibody, as well as in the following brain cell lines: D341 Med, SH-SY5Y, U-138 MG, U-87 MG. Glial tumor samples from up to 12 patients are also represented. In addition to the IHC images, each antibody is used for immunofluorescence (IF) staining in U-251 MG for subcellular localization information. A representation of products is provided in the Materials table below.

Neural Lineage Markers

Secretagogin is a newly discovered EF-hand calcium binding protein strongly expressed in the mouse olfactory bulb. Here visualized using the Anti-SCGN antibody (HPA006641).

Materials

     

 Reference

  1. Stem cells: attributes, cycles, spirals, pitfalls and uncertainties - Potten et al., Development, v110, 1001-1020 (1990)
  2. Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors – Yamanaka et al., Cell, Volume 126, Issue 4, 25 August 2006, Pages 663–676
  3. Rapid induction and long-term self-renewal of primitive neural precursors from human embryonic stem cells by small molecule inhibitors - Ding el al., PNAS, v108, 8299-8304 (2011)
  4. Small Molecules Greatly Improve Conversion of Human-Induced Pluripotent Stem Cells to the Neuronal Lineage - Mak et al, Stem Cells International (2012)
  5. http://stemcells.nih.gov/info/2001report/Pages/2001report.aspx
  6. Querkopf is a key marker of self-renewal and multipotency of adult neural stem cells - Sheikh et al., Journal of Cell Science. v125, 295-309 (2011)
  7. Querkopf, a MYST family histone acetyltransferases, is required for normal cerebral cortex development - Thomas et al., Development. v127, 2537-2548 (2000)
  8. Embryonic Stem Cell-Derived Neural Stem Cells Fuse with Microglia and mature neurons - Cusulin et al, Stem Cells, v30, 2657- 2671 (2012)
  9. Mulder J, Spence L, Tortoriello G, Dinieri JA, Uhlén M, Shui B, Kotlikoff MI, Yanagawa Y, Aujard F, Hökfelt T, Hurd YL, Harkany T. Secretagonin is a Ca2+ binding protein identifying prospective extended amygdala neurons in the developing mammalian telencephalon. Eur J Neurosci. 2010 Jun;31(12):2166-77 Epub 2010 Jun 7
  10. Mulder J, Zilberter M, Spence L, Tortoriello G, Uhlén M, Yanagawa Y, Aujard F, Hökfelt T, Harkany T. Secretagonin is a Ca2+ binding protein specifying subpopulations of telencephalic neurons. Proc Natl Acad Sci U S A. 2009 Dec 29;106(52):22492-7 Epub 2009 Dec 16.

 

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