Culture bag systems for clinical applications of adult human neural crest-derived stem cells

Greiner J, Grunwald L-M, Müller J, Sudhoff H, Widera D, Kaltschmidt C, Kaltschmidt B (2014)
Stem Cell Research & Therapy 5(2).

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Abstract
Introduction Facing the challenging treatment of neurodegenerative diseases as well as complex craniofacial injuries such as those common after cancer therapy, the field of regenerative medicine increasingly relies on stem cell transplantation strategies. Here, neural crest-derived stem cells (NCSCs) offer many promising applications, although scale up of clinical-grade processes prior to potential transplantations is currently limiting. In this study, we aimed to establish a clinical-grade, cost-reducing cultivation system for NCSCs isolated from the adult human nose using cGMP-grade Afc-FEP bags. Methods We cultivated human neural crest-derived stem cells from inferior turbinate (ITSCs) in a cell culture bag system using Afc-FEP bags in human blood plasma-supplemented medium. Investigations of viability, proliferation and expression profile of bag-cultured ITSCs were followed by DNA-content and telomerase activity determination. Cultivated ITSCs were introduced to directed in vitro differentiation assays to assess their potential for mesodermal and ectodermal differentiation. Mesodermal differentiation was determined using an enzyme activity assay (alkaline phosphatase, ALP), respective stainings (Alizarin Red S, Von Kossa and Oil Red O), and RT-PCR, while immunocytochemistry and synaptic vesicle recycling were applied to assay neuroectodermal differentiation of ITSCs. Results When cultivated within Afc-FEP bags, ITSCs grew three-dimensionally in a human blood plasma-derived matrix, thereby showing unchanged morphology, proliferation capability, viability and expression profile in comparison to three dimensionally-cultured ITSCs growing in standard cell culture plastics. Genetic stability of bag-cultured ITSCs was further accompanied by unchanged telomerase activity. Importantly, ITSCs retained their potential to differentiate into mesodermal cell types, particularly including ALP-active, Alizarin Red S-, and Von Kossa-positive osteogenic cell types, as well as adipocytes positive in Oil Red O assays. Bag culture further did not affect the potential of ITSCs to undergo differentiation into neuroectodermal cell types coexpressing β-III-tubulin and MAP2 and exhibiting the capability for synaptic vesicle recycling. Conclusions Here, we report for the first time the successful cultivation of human NCSCs within cGMP-grade Afc-FEP bags using a human blood plasma-supplemented medium. Our findings particularly demonstrate the unchanged differentiation capability and genetic stability of the cultivated NCSCs, suggesting the great potential of this culture system for future medical applications in the field of regenerative medicine.
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Greiner J, Grunwald L-M, Müller J, et al. Culture bag systems for clinical applications of adult human neural crest-derived stem cells. Stem Cell Research & Therapy. 2014;5(2).
Greiner, J., Grunwald, L. - M., Müller, J., Sudhoff, H., Widera, D., Kaltschmidt, C., & Kaltschmidt, B. (2014). Culture bag systems for clinical applications of adult human neural crest-derived stem cells. Stem Cell Research & Therapy, 5(2).
Greiner, J., Grunwald, L. - M., Müller, J., Sudhoff, H., Widera, D., Kaltschmidt, C., and Kaltschmidt, B. (2014). Culture bag systems for clinical applications of adult human neural crest-derived stem cells. Stem Cell Research & Therapy 5.
Greiner, J., et al., 2014. Culture bag systems for clinical applications of adult human neural crest-derived stem cells. Stem Cell Research & Therapy, 5(2).
J. Greiner, et al., “Culture bag systems for clinical applications of adult human neural crest-derived stem cells”, Stem Cell Research & Therapy, vol. 5, 2014.
Greiner, J., Grunwald, L.-M., Müller, J., Sudhoff, H., Widera, D., Kaltschmidt, C., Kaltschmidt, B.: Culture bag systems for clinical applications of adult human neural crest-derived stem cells. Stem Cell Research & Therapy. 5, (2014).
Greiner, Johannes, Grunwald, Lena-Marie, Müller, Janine, Sudhoff, Holger, Widera, Darius, Kaltschmidt, Christian, and Kaltschmidt, Barbara. “Culture bag systems for clinical applications of adult human neural crest-derived stem cells”. Stem Cell Research & Therapy 5.2 (2014).
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33 References

Data provided by Europe PubMed Central.

A highly enriched niche of precursor cells with neuronal and glial potential within the hair follicle dermal papilla of adult skin.
Hunt DP, Morris PN, Sterling J, Anderson JA, Joannides A, Jahoda C, Compston A, Chandran S., Stem Cells 26(1), 2008
PMID: 17901404
Human epidermal neural crest stem cells (hEPI-NCSC)--characterization and directed differentiation into osteocytes and melanocytes.
Clewes O, Narytnyk A, Gillinder KR, Loughney AD, Murdoch AP, Sieber-Blum M., Stem Cell Rev 7(4), 2011
PMID: 21455606
The human nose harbors a niche of olfactory ectomesenchymal stem cells displaying neurogenic and osteogenic properties.
Delorme B, Nivet E, Gaillard J, Haupl T, Ringe J, Deveze A, Magnan J, Sohier J, Khrestchatisky M, Roman FS, Charbord P, Sensebe L, Layrolle P, Feron F., Stem Cells Dev. 19(6), 2010
PMID: 19905894
Adult craniofacial stem cells: sources and relation to the neural crest.
Kaltschmidt B, Kaltschmidt C, Widera D., Stem Cell Rev 8(3), 2012
PMID: 22170630
Neurogenesis in the adult human hippocampus.
Eriksson PS, Perfilieva E, Bjork-Eriksson T, Alborn AM, Nordborg C, Peterson DA, Gage FH., Nat. Med. 4(11), 1998
PMID: 9809557
[Contamination of bovine fetal serum with bovine viral diarrhea virus]
Zabal O, Kobrak AL, Lager IA, Schudel AA, Weber EL., Rev. Argent. Microbiol. 32(1), 2000
PMID: 10785940

AUTHOR UNKNOWN, 2013
Large-scale generation of natural killer lymphocytes for clinical application.
Luhm J, Brand JM, Koritke P, Hoppner M, Kirchner H, Frohn C., J. Hematother. Stem Cell Res. 11(4), 2002
PMID: 12201953

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