Version 1
: Received: 25 May 2020 / Approved: 26 May 2020 / Online: 26 May 2020 (08:37:48 CEST)
Version 2
: Received: 1 November 2020 / Approved: 2 November 2020 / Online: 2 November 2020 (11:35:16 CET)
How to cite:
Grimm, W.; Didenko5, N.; Dhingra, K.; Dolgalev, A.; Enukashvily, N.; Fritsch, T.; Giesenhagen5, B.; Ivolgin, D.; Vukovic, M.A. Isolation of Neural Crest-derived Stem Cells for Therapeutic Use in Regenerative Periodontology. Preprints2020, 2020050427. https://doi.org/10.20944/preprints202005.0427.v1
Grimm, W.; Didenko5, N.; Dhingra, K.; Dolgalev, A.; Enukashvily, N.; Fritsch, T.; Giesenhagen5, B.; Ivolgin, D.; Vukovic, M.A. Isolation of Neural Crest-derived Stem Cells for Therapeutic Use in Regenerative Periodontology. Preprints 2020, 2020050427. https://doi.org/10.20944/preprints202005.0427.v1
Grimm, W.; Didenko5, N.; Dhingra, K.; Dolgalev, A.; Enukashvily, N.; Fritsch, T.; Giesenhagen5, B.; Ivolgin, D.; Vukovic, M.A. Isolation of Neural Crest-derived Stem Cells for Therapeutic Use in Regenerative Periodontology. Preprints2020, 2020050427. https://doi.org/10.20944/preprints202005.0427.v1
APA Style
Grimm, W., Didenko5, N., Dhingra, K., Dolgalev, A., Enukashvily, N., Fritsch, T., Giesenhagen5, B., Ivolgin, D., & Vukovic, M.A. (2020). Isolation of Neural Crest-derived Stem Cells for Therapeutic Use in Regenerative Periodontology. Preprints. https://doi.org/10.20944/preprints202005.0427.v1
Chicago/Turabian Style
Grimm, W., Dmitriy Ivolgin and Marco Alexander Vukovic. 2020 "Isolation of Neural Crest-derived Stem Cells for Therapeutic Use in Regenerative Periodontology" Preprints. https://doi.org/10.20944/preprints202005.0427.v1
Abstract
Periodontitis is microbial infection affecting periodontium, the tooth supporting structure and affects >743 million people worldwide. Neural crest-derived stem cells (NCSCs) hold the promise to regenerate the damaged periodontium. These cells have been identified within adult adipose tissue, periodontal ligament, and palatal tissue. Typical enzymatic isolation protocols are expensive, time consuming and often not clinically compliant. Enzyme-free, mechanical dissociation has been suggested as an alternative method of generating cell suspensions required for cell separation and subsequent expansion ex vivo. In our study, samples of rat skeletal muscle tissue were used to appraise the suitability of a novel mincing method of mechanical dissociation against enzymatic digestion with collagenase and dispase. Skeletal muscle is readily available and has been shown to contain NCSC populations. We used a Rigenera-Human Brain Wave® prototype mincer to produce a suspension of NCSCs. We have compared the resulting cell cultures produced via mechanical dissociation and enzymatic dissociation, producing single cell suspensions suitable for Magnetic Cell Sorting (MACs) and Fluorescence-activated cell sorting (FACS). Despite the Countess Automated Cytometry data demonstrating that cell suspensions produced by mechanical dissociation (n=24) contain on average 26.8 times as many viable cells as enzymatic cell suspensions (n=18), enzymatic suspensions produced more successful cell cultures. Spheroids and subsequently adherent cells formed from 4 enzymatic cell suspensions (44.4%) vs. 1 mechanical cell suspension (8.3%). Enzymatic digestion protocols formed spheroids faster and more plentifully than mechanical cell suspensions. Adherent cells and spheroids isolated via both methods appear morphologically similarly to NCSCs from our previous studies.
Medicine and Pharmacology, Dentistry and Oral Surgery
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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