Dimethyl sulfoxide-free cryopreservation solutions for hematopoietic stem cell grafts

Kaushal, Richa; Jahan, Suria; McGregor, Chelsea; Pineault, Nicolas

doi:10.1016/j.jcyt.2021.09.002

Cited by 11 publications

(5 citation statements)

References 42 publications

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“…In fact, studies have demonstrated high post-thaw cell viability in cells cryopreserved using non-DMSO cryoprotectant compared with DMSO-cryopreserving cells. 47 , 48 In this study, we have cryopreserved the dissociated TG neurons using both DMSO and DMSO-free cryoprotectants (CryoScarless), respectively, using our optimized protocol described above. However, no cells were found attached in the CryoScarless preserved group (data not shown), suggesting that CryoScarless might not be suitable in the cryopreservation of neuronal cells.…”

Section: Discussionmentioning

confidence: 99%

Culture of Primary Neurons from Dissociated and Cryopreserved Mouse Trigeminal Ganglion

Lin

Lee

Chen

et al. 2023

Tissue Engineering Part C: Methods

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Corneal nerves originate from the ophthalmic branch of the trigeminal nerve, which enters the cornea at the limbus radially from all directions toward the central cornea. The cell bodies of the sensory neurons of trigeminal nerve are located in the trigeminal ganglion (TG), while the axons are extended into the three divisions, including ophthalmic branch that supplies corneal nerves. Study of primary neuronal cultures established from the TG fibers can therefore provide a knowledge basis for corneal nerve biology and potentially be developed as an in vitro platform for drug testing. However, setting up primary neuron cultures from animal TG has been dubious with inconsistency among laboratories due to a lack of efficient isolation protocol, resulting in low yield and heterogenous cultures. In this study, we used a combined enzymatic digestion with collagenase and TrypLE to dissociate mouse TG while preserving nerve cell viability. A subsequent discontinuous Percoll density gradient followed by mitotic inhibitor treatment effectively diminished the contamination of non-neuronal cells. Using this method, we reproducibly generated high yield and homogenous primary TG neuron cultures. Similar efficiency of nerve cell isolation and culture was further obtained for TG tissue cryopreserved for short (1 week) and long duration (3 months), compared to freshly isolated tissues. In conclusion, this optimized protocol shows a promising potential to standardize TG nerve culture and generate a high-quality corneal nerve model for drug testing and neurotoxicity studies. Impact statement Isolation of neurons from trigeminal ganglion (TG) has been used for deeper insights on corneal nerve biology. However, the lack of standardized isolation protocols may be the attributes of the heterogenous culture with inconsistent results among laboratories. In this study, we investigated an optimized protocol for TG neuron culture with good efficiency. Furthermore, compared with freshly prepared TG culture, the culture prepared with cryopreserved TG tissues or neurons showed similar morphology with preserved neuronal marker expression. This study highlights the promising potential of using optimized protocol as a standardized isolation method as in vitro corneal nerve model for corneal nerve studies.

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Section: Discussionmentioning

confidence: 99%

Culture of Primary Neurons from Dissociated and Cryopreserved Mouse Trigeminal Ganglion

Lin

Lee

Chen

et al. 2023

Tissue Engineering Part C: Methods

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“…Finally, reducing the concentration of DMSO in the cryoprotective mixture to 5% effectively minimizes the amount of infused DMSO, resulting in a lower incidence of adverse events, without affecting hematopoietic recovery [ 39 ]. In the next future, new DMSO-free freezing media could replace DMSO and provide adequate cryoprotection to HSC grafts limiting toxicities to cells and recipients [ 40 ].…”

Section: Cryopreservation Of Hematopoietic Stem Cell Graftsmentioning

confidence: 99%

Pros and Cons of Cryopreserving Allogeneic Stem Cell Products

Valentini,

Pellegrino,

Teofili

2024

Cells

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The COVID-19 pandemic has precipitously changed the practice of transplanting fresh allografts. The safety measures adopted during the pandemic prompted the near-universal graft cryopreservation. However, the influence of cryopreserving allogeneic grafts on long-term transplant outcomes has emerged only in the most recent literature. In this review, the basic principles of cell cryopreservation are revised and the effects of cryopreservation on the different graft components are carefully reexamined. Finally, a literature revision on studies comparing transplant outcomes in patients receiving cryopreserved and fresh grafts is illustrated.

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“…Имеются исследования, показывающие, что использование ДМСО снижает экспрессию факторов транскрипции, таких как Oct-4, Sox-2, Nanog и Rex-1, специфичных для эмбриональных стволовых клеток человека [48,49], а также влияние на эпигенетическое состояние клеток [50]. Предполагается, что добавление новых криопротекторов позволит улучшить жизнеспособность клеток после оттаивания и повысит успешность трансплантации [51].…”

Section: методология поискаunclassified

Cryostorage of peripheral blood hematopoietic stem cells in transplantology: current status and prospects

Kit,

Gnennaya,

Filippova

et al. 2023

Cardiovasc Ther Prev

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Peripheral blood hematopoietic stem cell (HSC) transplantation is a well-established procedure for the treatment of hematological, cancer and autoimmune diseases. In cancer patients, HSC transplantation allows the use of high-dose cytotoxic drugs in combination with radiation therapy during treatment, which provides a pronounced antitumor effect. The hematological toxicity of such treatment is eliminated by the sequential introduction of stem cells, which contribute to hematopoiesis restoration. Before transplantation, peripheral blood HSCs are subjected to collection and cryopreservation for further storage. An important requirement for cryopreservation is viable HSCs responsible for hematopoietic restoration. The aim of the review was to analyze the literature devoted to the influence of various methods of cryopreservation of human peripheral blood HSCs on the preservation of cell viability after thawing, as well as the development of adverse events in patients. Issues related to the use of various cryoprotectants, as well as methods for storing HSC grafts, are considered. The presented data indicate the need for further study of the effect of cryoprotectants on the human body and the cellular composition of the graft and improvement of protocols for HSC cryopreservation.

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Dimethyl sulfoxide-free cryopreservation solutions for hematopoietic stem cell grafts

Cited by 11 publications

References 42 publications

Culture of Primary Neurons from Dissociated and Cryopreserved Mouse Trigeminal Ganglion

Culture of Primary Neurons from Dissociated and Cryopreserved Mouse Trigeminal Ganglion

Pros and Cons of Cryopreserving Allogeneic Stem Cell Products

Cryostorage of peripheral blood hematopoietic stem cells in transplantology: current status and prospects

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