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StemBeads® Qkine GDNF
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StemBeads® Qkine GDNF

StemBeads® Qkine GDNF is a patented growth factor supplement that offers a novel way to culture cells with native GDNF more efficiently, with greater control and fewer medium changes. StemBeads® Qkine GDNF has been tested on neural progenitor cell (NPC) media, including neural progenitor expansion medium (NPEM), with enhanced cellular profiles.

StemBeads® revolutionary controlled-release protein technology combined with Qkine high purity and bioactivity animal origin-free growth factors and cytokines stabilizes growth factor levels for increased cell culture consistency and reproducibility.

 

Our robust animal origin-free manufacturing platform, along with rigorous quality control procedures, ensures exceptional bioactivity and consistent performance from lot to lot, guaranteeing outstanding performance in your applications.

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StemBeads® Qkine GDNF is a patented growth factor supplement that offers a novel way to culture cells with native Glial Derived Neurotrophic Factor more efficiently, with greater control and fewer medium changes. GDNF is key for supporting neural progenitor cells (NPCs) and neuronal differentiation. StemBeads® Qkine GDNF are microbeads composed of an FDA approved, biodegradable PLGA polymer that is loaded with Qkine recombinant human GDNF (Qk051) and release the encapsulated protein at a constant rate. Controlled delivery and stable levels overcome the 34 minute half-life (Figure 1) of GDNF and improve cell cultures while saving researchers valuable time and resources.

StemBeads are a revolutionary controlled-release growth factor technology. The StemBeads product is currently used for improved cell culture in more than 100 scientific laboratories globally. StemBeads technology encapsulates a growth factor in a PLGA hydrogel that stabilizes the protein and releases it steadily over several days. With growth factor levels stabilized, spontaneous differentiation is reduced, pluripotency is increased, and feeding schedules are reduced to only 2-3 times per week. StemBeads are added to your medium to achieve a desired growth factor release, sustained over several days. Following, the beads are washed from the culture, the medium is replaced, and the process repeated as required.

Glial cell line-derived neurotrophic factor (GDNF) is a neurotrophic factor and a member of the GDNF family of ligands (GFL) which include neurturin, artemin, and persephin [1,2]. GFLs regulate several biological processes including cell survival, cell proliferation, cell differentiation, cell migration, and neurite outgrowth [1,3,4]. GDNF protein plays a crucial role in the development, growth, and survival of neurons in particular midbrain dopaminergic neurons [5]. It promotes the axon growth and innervation of dopamine neurons. It also facilitates the differentiation of neural progenitors to astrocytes and is involved in neuroprotection[6,7]. In addition, it is involved in kidney development and spermatogenesis [8,9].

In cell culture, GDNF supports the survival and growth of neurons and astrocytes. It is used in combination with other growth factors to differentiate human induced pluripotent stem cell-derived neural progenitors into neurons and astrocytes. GDNF is used for the differentiation, survival, and maturation of dopaminergic neurons with brain-derived neurotrophic factor (BDNF), FGF-8a/FGF-8b, and Shh [10-13]. Glutamatergic and GABAergic neurons are obtained with combinations of BDNF, GDNF, NT-3, or insulin-like growth factor 1 (IGF-1) [14-16]. Also, the culture of cortical organoids requires BDNF and GDNF for maturation [17,18]. Finally, it can be used to generate and maintain astrocytes along with ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF) [19-21].

[1] Airaksinen, M. S. & Saarma, M. The GDNF family: Signalling, biological functions and therapeutic value. Nat. Rev. Neurosci. 3, 383–394 (2002). doi: 10.1038/nrn812

[2] von dem Bussche, M. & Tuszynski, M. H. Growth Factors: Neuronal Atrophy. in Encyclopedia of Neuroscience (ed. Squire, L. R.) 987–992 (Academic Press, 2009). doi:10.1016/B978-008045046-9.00143-1.

[3] Airaksinen, M. S., Titievsky, A. & Saarma, M. GDNF Family Neurotrophic Factor Signaling: Four Masters, One Servant? Mol. Cell. Neurosci. 13, 313–325 (1999). doi: 10.1006/mcne.1999.0754

[4] Baloh, R. H., Enomoto, H., Johnson, E. M. & Milbrandt, J. The GDNF family ligands and receptors — implications for neural development. Curr. Opin. Neurobiol. 10, 103–110 (2000). doi: 10.1016/s0959-4388(99)00048-3

[5] Akerud, P., Alberch, J., Eketjäll, S., Wagner, J. & Arenas, E. Differential effects of glial cell line-derived neurotrophic factor and neurturin on developing and adult substantia nigra dopaminergic neurons. J. Neurochem. 73, 70–78 (1999). doi: 10.1046/j.1471-4159.1999.0730070.x

[6] Parsadanian, A., Pan, Y., Li, W., Myckatyn, T. M. & Brakefield, D. Astrocyte-derived transgene GDNF promotes complete and long-term survival of adult facial motoneurons following avulsion and differentially regulates the expression of transcription factors of AP-1 and ATF/CREB families. Exp. Neurol. 200, 26–37 (2006). doi: 10.1016/j.expneurol.2006.01.014

[7] Boku, S. et al. GDNF facilitates differentiation of the adult dentate gyrus-derived neural precursor cells into astrocytes via STAT3. Biochem. Biophys. Res. Commun. 434, 779–784 (2013). doi.org/10.1016/j.bbrc.2013.04.011

[8] Costantini, F. & Shakya, R. GDNF/Ret signaling and the development of the kidney. BioEssays News Rev. Mol. Cell. Dev. Biol. 28, 117–127 (2006). doi.org/10.1002/bies.20357

[9] Hofmann, M.-C. Gdnf signaling pathways within the mammalian spermatogonial stem cell niche. Mol. Cell. Endocrinol. 288, 95–103 (2008). doi: 10.1016/j.mce.2008.04.012

[10] Granholm, A.-C. et al. Glial Cell Line-Derived Neurotrophic Factor Is Essential for Postnatal Survival of Midbrain Dopamine Neurons. J. Neurosci. 20, 3182–3190 (2000). doi.org/10.1523/JNEUROSCI.20-09-03182.2000

[11] Perrier, A. L. et al. Derivation of midbrain dopamine neurons from human embryonic stem cells. Proc. Natl. Acad. Sci. U. S. A. 101, 12543–12548 (2004). doi.org/10.1073/pnas.0404700101

[12] Brennand, K. et al. Modeling schizophrenia using hiPSC neurons. Nature 473, 221–225 (2011). doi: 10.1038/nature09915

[13] Kriks, S. et al. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease. Nature 480, 547–551 (2011). doi.org/10.1038/nature10648

[14] Fernandopulle, M. S. et al. Transcription-factor mediated differentiation of human iPSCs into neurons. Curr. Protoc. Cell Biol. 79, e51 (2018). doi: 10.1002/cpcb.51

[15] Li, X.-J. et al. Specification of motoneurons from human embryonic stem cells. Nat. Biotechnol. 23, 215–221 (2005). doi: 10.1038/nbt1063

[16] Lin, L., Yuan, J., Sander, B. & Golas, M. M. In Vitro Differentiation of Human Neural Progenitor Cells Into Striatal GABAergic Neurons. Stem Cells Transl. Med. 4, 775–788 (2015). doi: 10.5966/sctm.2014-0083

[17] McComish, S. F. & Caldwell, M. A. Generation of defined neural populations from pluripotent stem cells. Philos. Trans. R. Soc. B Biol. Sci. 373, 20170214 (2018). doi: 10.1098/rstb.2017.0214

[18] Jacob, F. et al. Human Pluripotent Stem Cell-Derived Neural Cells and Brain Organoids Reveal SARS-CoV-2 Neurotropism Predominates in Choroid Plexus Epithelium. Cell Stem Cell 27, 937-950.e9 (2020). doi: 10.1016/j.stem.2020.09.016

[19] Krencik, R., Weick, J. P., Liu, Y., Zhang, Z. & Zhang, S.-C. Specification of Transplantable Astroglial Subtypes from Human Pluripotent Stem Cells. Nat. Biotechnol. 29, 528–534 (2011). doi: 10.1038/nbt.1877

[20] Serio, A. et al. Astrocyte pathology and the absence of non-cell autonomy in an induced pluripotent stem cell model of TDP-43 proteinopathy. Proc. Natl. Acad. Sci. U. S. A. 110, 4697–4702 (2013). doi.org/10.1073/pnas.1300398110

[21] Gunhanlar, N. et al. A simplified protocol for differentiation of electrophysiologically mature neuronal networks from human induced pluripotent stem cells. Mol. Psychiatry 23, 1336–1344 (2018). doi.org/10.1038/mp.2017.56

Additional information

FAQ

StemBeads® can be easily removed prior to analysis by washing 1-2 times with PBS, DMEM, F12 or any other basal medium preferred. For suspension cultures, there are a variety of strainer pore sizes to allow for separation. Cells cultured with StemBeads® have been used in a variety of downstream applications, including but not limited to: immunofluorescence, FACS, RNA/DNA extraction, nucleofection, differentiation etc.

The protein concentration can be adjusted based on the number of StemBeads® that you use. The release rate has a linear correlation with the amount of StemBeads® used. For example, 8 µl of StemBeads® per ml of media releases at a concentration of 10 ng/ml. However, if you desire half of that concentration, such as 5 ng/ml, then adjust the concentration to 4 µl of StemBeads® per ml media to achieve the concentration of 5 ng/ml.

With growth factor levels stabilized, spontaneous differentiation is reduced, pluripotency is increased, and feeding schedules are reduced to only 2-3 times per week. StemBeads are added to your medium to achieve a desired growth factor release, sustained over several days. Following, the beads are washed from the culture, the medium is replaced, and the process repeated as required.

No, this is not a problem or indication of cell quality. Extensive testing has been performed to access the cell quality when using StemBeads®. All phenotypic and functional assays have shown cell quality as good or better than current feeding regimens. For best assessment of cells, wash cells once prior to observation.

GDNF protein plays a crucial role in the development, growth, and survival of midbrain dopaminergic neurons.

GDNF is mainly released by glial cells such as astrocytes, Schwann cells, and satellite cells.

GDNF is used as a growth factor which is supplemented in cell culture media to promote the differentiation, growth, and maturation of dopaminergic neurons.

Qkine products are intended solely for research purposes and ex vivo cell manufacturing. Not for therapeutic or diagnostic use.