Optimization of quality parameters for human thymic cell samples stored in liquid nitrogen

Shichkin, Valentin P.; Gorbach, Oleksandr; Zuieva, O. A.; Martsenyuk, Olga P.

doi:10.15761/tit.1000244

Cited by 4 publications

(12 citation statements)

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“…These thymus-derived cell cultures contain both TESC and their differentiated progenies as well as MSC and fibroblasts. Importantly, some of the cultured cells also retain the capacity to contribute to thymic stromal-epithelial networks where they exhibit a normal thymic function in terms of T cell differentiation from CD34+ hematopoietic stem cells [39,66] and Antica, unpublished data. Thus, the current culture conditions, which are optimized for epithelial stem cells can be used as a starting point to define optimal conditions for an effective support of the human TESC growth.…”

Section: Thymic Epithelial Cell Linesmentioning

confidence: 97%

“…In mice they express Plet1, Ly51, and EpCAM (CD326) surface proteins [38••]. CD326 is also expressed in the fetal human thymus, and therefore, in combination with Foxn1 expression could be used to identify the human TEPC/TESC population(s) [26,39]. Recently, in the course of the ThymiStem project funded by European Union, Prof. Antica's research group has detected epithelial precursors also from human thymus by using the stem cell ability to form spheres when cultured in non-adherent conditions in vitro (manuscript in preparation).…”

Section: Thymus Cell Architecture and Thymic Epithelial Cellsmentioning

confidence: 99%

“…While cells grown under these conditions can contribute to epithelial networks, they do so at low efficiency and further optimization for an increase of the functional stem cell frequency is required to develop clinically useful lines. Functional cultures of thymic stromal-epithelial stem cells can be derived also in low-adhesive conditions as cultures of thymospheres [15,40] or in adhesive conditions as cultures of thymic explants and monolayer cultures [39,65,66]. These thymus-derived cell cultures contain both TESC and their differentiated progenies as well as MSC and fibroblasts.…”

Section: Thymic Epithelial Cell Linesmentioning

confidence: 99%

“…Currently, various freezing media designed specifically for cryopreservation of stem cells are available from different manufacturers, and some of these were applied for the cryopreservation and the long-term storage of the human thymic tissue in liquid nitrogen [65,87]. A specific investigation concerning cryopreservation of postnatal thymic tissue in a wide range of cryoprotective conditions has been implemented by Prof. Shichkin's research group during the ThymiStem project [39,65]. In this study, the influence of a number of cryoprotective media with either penetrating (DMSO, glycerol) or non-penetrating (dextran-40, sucrose, hydroxyethyl starch) components was evaluated, and compared to the commercial GMP manufactured cryoprotective medium Stem-CellBanker (AMS Biotechnology, UK).…”

Section: Thymic Tissue Cryopreservationmentioning

confidence: 99%

“…Stem-CellBanker is serum-free and DMSO-containing medium, and it was created specifically for stem cell storage. This study indicated that for either cell suspensions or thymic fragments, the best combination for long-term storage was DMSO and dextran-40 (CPM-7) as judged by the CD326 + epithelial cells' viability and formation of a stromal-epithelial cell monolayer after thawing [39,65]. This cryoprotective medium could be further optimized specifically for human TESC in comparison with the set of available commercial media.…”

Section: Thymic Tissue Cryopreservationmentioning

confidence: 99%

See 4 more Smart Citations

Thymus Regeneration and Future Challenges

Shichkin¹,

Antica

2020

Stem Cell Rev and Rep

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Thymus regenerative therapy implementation is severely obstructed by the limited number and expansion capacity in vitro of tissue-specific thymic epithelial stem cells (TESC). Current solutions are mostly based on growth factors that can drive differentiation of pluripotent stem cells toward tissue-specific TESC. Target-specific small chemical compounds represent an alternative solution that could induce and support the clonal expansion of TESC and reversibly block their differentiation into mature cells. These compounds could be used both in the composition of culture media designed for TESC expansion in vitro, and in drugs development for thymic regeneration in vivo. It should allow reaching the ultimate objective-autologous thymic tissue regeneration in paediatric patients who had their thymus removed in the course of cardiac surgery.

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Section: Thymic Epithelial Cell Linesmentioning

confidence: 97%

Section: Thymus Cell Architecture and Thymic Epithelial Cellsmentioning

confidence: 99%

Section: Thymic Epithelial Cell Linesmentioning

confidence: 99%

Section: Thymic Tissue Cryopreservationmentioning

confidence: 99%

Section: Thymic Tissue Cryopreservationmentioning

confidence: 99%

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Thymus Regeneration and Future Challenges

Shichkin¹,

Antica

2020

Stem Cell Rev and Rep

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Stem Cells of the Thymus

Shichkin

2021

Stem Cells: From Potential to Promise

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A model for preservation of thymocyte-depleted thymus

Dias

Damaceno-Rodrigues

Gimenez

et al. 2023

Braz J Med Biol Res

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DiGeorge syndrome is a disorder caused by a microdeletion on the long arm of chromosome 22. Approximately 1% of patients diagnosed with DiGeorge syndrome may have an absence of a functional thymus, which characterizes the complete form of the syndrome. These patients require urgent treatment to reconstitute T cell immunity. Thymus transplantation is a promising investigational procedure for reconstitution of thymic function in infants with congenital athymia. Here, we demonstrate a possible optimization of the preparation of thymus slices for transplantation through prior depletion of thymocytes and leukocyte cell lineages followed by cryopreservation with cryoprotective media (5% dextran FP 40, 5% Me2SO, and 5% FBS) while preserving tissue architecture. Thymus fragments were stored in liquid nitrogen at -196°C for 30 days or one year. The tissue architecture of the fragments was preserved, including the distinction between medullary thymic epithelial cells (TECs), cortical TECs, and Hassall bodies. Moreover, depleted thymus fragments cryopreserved for one year were recolonized by intrathymic injections of 3×10 6 thymocytes per mL, demonstrating the capability of these fragments to support T cell development. Thus, this technique opens up the possibility of freezing and storing large volumes of thymus tissue for immediate transplantation into patients with DiGeorge syndrome or atypical (Omenn-like) phenotype.

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Optimization of quality parameters for human thymic cell samples stored in liquid nitrogen

Cited by 4 publications

References 0 publications

Thymus Regeneration and Future Challenges

Thymus Regeneration and Future Challenges

Stem Cells of the Thymus

A model for preservation of thymocyte-depleted thymus

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