StemPro NSC SFM—Serum-Free Human Neural Stem Cell Culture Medium

• Superior expansion of neural stem cells (NSCs) derived from either embryonic stem cells or from fetal tissue (see Figure 2)
• Versatility to support long-term growth and expansion of both adherent and neurosphere suspension cultures
• Maintain normal NSC multipotency and phenotype/karyotype
• NSCs grown in StemPro NSC SFM maintain the potential to differentiate to physiologically active neurons and glial cells
• Better batch-to-batch consistency, with each lot produced under cGMP and qualified using an hNSC performance assay
• No or little adaptation required from serum-supplemented medium

Nestin

Sox2/Oct4

Ki67

Figure 1.  Phenotype marker expression of neural stem cells cultured in StemPro NSC SFM.  Phenotype marker expression of NSC after culture in StemPro NSC SFM through passage 17.  NSCs expressed normal phenotypic markers (Nestin, Sox2) and proliferation marker (Ki67).  There was no sign of remnant hESC (Oct4).  The inset image in each panel shows the staining pattern given by the nuclear stain DAPI.

StemPro NSC SFM delivers superior hNSC expansion and versatility for both adherent and neurosphere suspension cultures.  As human NSCs comprise a very small fraction of the total CNS cell population, expansion is critical to generate sufficient cells to study differentiation pathways and explore the downstream clinical applications of human NSCs.  hNSCs can be passaged only a limited number of times before exhibiting reduced proliferation and differentiation potential.  Maximizing the total hNSC yield per passage is therefore essential (Figure 2).

Figure 2.  Superior human NSC expansion is achieved using StemPro NSC SFM compared with competitor serum-free NSC media. StemPro NSC SFM demonstrates superior cell expansion capacity compared with standard N2-supplemented and competitor neural stem cell media formulations. Proliferation of hNSCs cultured in Invitrogen StemPro NSC SFM, competitor SCT medium, Sigma medium, and N2 supplemented medium was measured. ESC-derived hNSCs were seeded at 1 x 10⁴ cells per well in CELLstart substrate–coated 96-well plates for 3 days in respective media. Indirect cell count was obtained with the CyQUANT proliferation assay kit (Cat. no. C35006), and data shows mean relative florescence units of stained cells (n=6).

(A) Adherent Culture

(B) Suspension Culture

Figure 3.  Stable proliferation of hNSCs in StemPro NSC SFM enables culturing of both adherent and suspension culture systems.  StemPro NSC SFM provides the flexibility to culture hNSCs for several passages, maintaining multipotent characteristics as either an (A) adherent or (B) neurosphere culture. hNSCs were derived from hESCs cultured in NSC SFM for 7 passages on CELLstart substrate.  Tertiary neurospheres were isolated from fetal tissue cultured in NSC SFM.

hNSCs are defined by the ability to differentiate to three distinct lineages-neurons, oligodendrocytes, and astrocytes.  StemPro NSC SFM delivers a robust, serum-free neural stem cell medium that maintains the multipotent differentiation capabilities of the stem cells and the ability to drive hNSCs down the desired lineage to meet specific experimental requirements (Figure 4).

Panel A

Panel B

Panel C

Figure 4.  Differentiation potential of hNSCs cultured in StemPro NSC SFM.  hNSCs were cultured in StemPro NSC SFM and were differentiated to neurons, and glial cells.  Shown above (A) Neurons were labeled with an anti-HuC/D antibody (green) and an anti-Dcx antibody (red).  (B) Cells with an oligodendrocyte lineage were labeled with an anti-GalC antibody (red).  Cell nuclei were labeled with DAPI (blue) and neurons were labeled with an anti-Dcx antibody (green).  (C) Cells with an astrocyte lineage were labeled using an anti-CD44 antibody (green).  Cell nuclei were labeled with DAPI (blue) and neurons were labeled with an anti-Dcx antibody (green).