Cryopreservation of Dental Stem Cells

Abstract

Nowadays, regenerative and reparative medicine has grown in popularity. Dental stem cells are easily accessible source of adult stem cells. They can be harvested by a tooth extraction or spontaneous deciduous tooth exfoliation. They have to be isolated, expanded and stored until time they would be needed for individual stem cell therapy. Cryopreservation is both a short-term and long-term storage of tissues or cells at sub-zero temperatures. There are several methods of cryopreservation requiring different technologies. The objective of this review is to compare them and highlight their advantages and disadvantages.

Dental stem cell banking: Techniques and protocols

“… Note : Arora et al (2009); Gronthos et al (2000); Hegde et al (2012); Kang et al (2019); Karamzadeh et al (2012); Lindemann et al (2014); J. Liu et al (2014); Mori et al (2012); Pilbauerová and Suchánek (2018); Rémy et al (2019); W. Zhang et al (2006); Zhu and Liang (2015). …”

“…Also a specific buffer providing nutrients and preventing the tissue from drying out, is required to ensure that the samples will remain viable (Arora et al, 2009). A chilled sterile vial containing a hypotonic phosphate‐buffered saline solution (PBS) are commonly used as a transfer buffer (Agarwal et al, 2016; Arora et al, 2009; Derakhshani et al, 2015; Hegde et al, 2012; Perry et al, 2008; Pilbauerová & Suchánek, 2018), however many companies provide commercial transport kits for transporting stem cells from the dental office to the laboratory (Table 3). Other reported collection/transport solutions were HypoThermosol (BioLife Solutions), MesenCult basal medium (StemCell Technologies), and basic medium (alpha minimal essential medium [α‐MEM] supplemented with 10% fetal bovine serum [FBS]) (Karamzadeh et al, 2012; Perry et al, 2008).…”

“…For DPSCs, the pulp chamber should be revealed by cutting the teeth around the cementum–enamel junction by using sterilized dental fissure burs (Gronthos et al, 2000; Karamzadeh et al, 2012; Pilbauerová & Suchánek, 2018). For SHED, since the pulp chamber is already exposed, the pulp tissue could be removed using sterile small forceps or a dental excavator (Arora et al, 2009).…”

“…For PDLSCs, the PDL needs to be gently separated from the middle part of the root surface and cut into small pieces (J. Liu et al, 2014; Wang et al, 2017). For all types of DSCs, both spontaneous outgrowth (explant method) and enzymatic dissociation methods were widely used (Hegde et al, 2012; Hollands et al, 2018; Pilbauerová & Suchánek, 2018; Zhu & Liang, 2015). Although enzymatic digestion seems more efficient, considering translation to clinical applications, the explant method seems to be the preferred approach (Hilkens et al, 2016).…”

Intracellular delivery of trehalose renders mesenchymal stromal cells viable and immunomodulatory competent after cryopreservation

“…Cryopreservation at sub-zero temperatures is the most widely used method for long-term storage of cells and tissues, which allows preservation of their vital functions upon thawing Dulugiac et al 2015 ; Marquez-Curtis et al 2015 ; Mitchell et al 2015 ; Pilbauerová et al 2018 ; Ramos et al 2014 ; Yong 2015 ). As the use of cell-based therapies advances, there is thus also the need for safer and more effective cryopreservation protocols for these cell products (Awan et al 2020 ; de Luca et al 2019 ; Hornberger et al 2019 ; Weng and Beauchesne 2020 ).…”