|StemPro NSC SFM is specifically formulated for serum-free growth and expansion of human neural stem cells (hNSCs). |
Superior NSC expansion and versatility with serum-free media
- 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
hNSCs maintain multipotency and normal phenotype when grown in StemPro NSC SFM
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
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).
StemPro NSC SFM provides the versatility to expand and maintain both adherent and neurosphere suspensions in culture
(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.
StemPro NSC SFM maintains the multipotent differentiation capabilities of hNSCs
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).
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).
|CELLstart defined, xeno-free cell culture substrate for stem cells|
Learn more about CELLstart substrate
- Wu YY, Mujtaba T, Rao MS. Isolation of stem and precursor cells from fetal tissue. Methods Mol Biol 2002;198:29-40.
- Bjorklund A, Lindvall O. Cell replacement therapies for central nervous system disorders. Nat Neurosci 2000;3(6):537-44.
- Gage FH. Mammalian neural stem cells. Science 2000;287(5457):1433-8.
- Studer L, Tabar V, McKay RD. Transplantation of expanded mesencephalic precursors leads to recovery in parkinsonian rats. Nat Neurosci 1998;1(4):290-5.
- Caldwell MA, He X, Wilkie N, et al. Growth factors regulate the survival and fate of cells derived from human neurospheres. Nat Biotechnol 2001;19(5):475-9.
- Jain M, Armstrong RJ, Tyers P, Barker RA, Rosser AE. GABAergic immunoreactivity is predominant in neurons derived from expanded human neural precursor cells in vitro. Exp Neurol 2003;182(1):113-23.
- Reubinoff BE, Itsykson P, Turetsky T, et al. Neural progenitors from human embryonic stem cells. Nat Biotechnol 2001;19(12):1134-40.
- Shin S, Mitalipova M, Noggle S, et al. Long-term proliferation of human embryonic stem cell-derived neuroepithelial cells using defined adherent culture conditions. Stem Cells 2006;24(1):125-38.
- Colombo E, Giannelli SG, Galli R, et al. Embryonic stem-derived versus somatic neural stem cells: a comparative analysis of their developmental potential and molecular phenotype. Stem Cells 2006;24(4):825-34.
- Watanabe K, Kamiya D, Nishiyama A, et al. Directed differentiation of telencephalic precursors from embryonic stem cells. Nat Neurosci 2005;8(3):288-96.
- Mayer-Proschel M, Kalyani AJ, Mujtaba T, Rao MS. Isolation of lineage-restricted neuronal precursors from multipotent neuroepithelial stem cells. Neuron 1997;19(4):773-85.
- Cai J, Wu Y, Mirua T, et al. Properties of a fetal multipotent neural stem cell (NEP cell). Dev Biol 2002;251(2):221-40.
- Uchida N, Buck DW, He D, et al. Direct isolation of human central nervous system stem cells. Proc Natl Acad Sci U S A 2000;97(26):14720-5.
- Gleeson JG, Lin PT, Flanagan LA, Walsh CA. Doublecortin is a microtubule-associated protein and is expressed widely by migrating neurons. Neuron 1999;23(2):257-71.
- Barami K, Iversen K, Furneaux H, Goldman SA. Hu protein as an early marker of neuronal phenotypic differentiation by subependymal zone cells of the adult songbird forebrain. J Neurobiol 1995;28(1):82-101.
- Zhang SC. Defining glial cells during CNS development. Nat Rev Neurosci 2001;2(11):840-3.
- Liu Y, Han SS, Wu Y, et al. CD44 expression identifies astrocyte-restricted precursor cells. Dev Biol 2004;276(1):31-46.
- Conti L, Pollard SM, Gorba T, et al. Niche-independent symmetrical self-renewal of a mammalian tissue stem cell. PLoS Biol 2005;3(9):e283.
- Wachs FP, Couillard-Despres S, Engelhardt M, et al. High efficacy of clonal growth and expansion of adult neural stem cells. Lab Invest 2003;83(7):949-62.
- Ostenfeld T, Svendsen CN. Requirement for neurogenesis to proceed through the division of neuronal progenitors following differentiation of epidermal growth factor and fibroblast growth factor-2-responsive human neural stem cells. Stem Cells 2004;22(5):798-811.