Healthy cloned offspring derived from freeze-dried somatic cells

Wakayama, Sayaka; Ito, Daiyu; Hayashi, Erika; Ishiuchi, Takashi; Wakayama, Teruhiko

doi:10.1038/s41467-022-31216-4

Cited by 14 publications

(8 citation statements)

References 53 publications

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“…Chromosomal analysis showed that the female clone had 39,X karyotype, indicating that the Y chromosome had been lost in the donor cell. The same situation has been observed in clones from ES cells [36], nuclear-transferred ES (ntES) cells derived from freeze-dried fibroblasts [45], and horse SCNT [46]. These findings show that SCNT could be a useful reproductive technique, especially for reviving endangered and extinct species.…”

Section: Selection Of Appropriate Cell Types For Mouse Scntsupporting

confidence: 62%

Mouse somatic cell nuclear transfer: What has changed and remained unchanged in 25 years

INOUE

2023

Journal of Reproduction and Development

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Somatic cell nuclear transfer (SCNT) is the only reproductive technology used to produce individuals from somatic cells by transferring them to enucleated oocytes. Although more than 25 years have passed since the first mammalian SCNT was reported in sheep, problems such as low birth rates and morphological abnormalities have persisted and limited its practical applications. The mouse is the ideal laboratory animal to unveil these questions due to its established reproductive technologies and extensive knowledge base of its genome and various strains. We investigated the causes of incomplete reprogramming after nuclear transfer of donor somatic cells and found that the loss of imprint in some placenta-specific imprinted genes could induce non-random SCNT abnormalities. By ameliorating aberrantly expressed imprinted genes, we succeeded in increasing the low birth rate and improving morphological abnormalities observed in SCNT fetuses. Furthermore, we sought appropriate mouse strains and cell types as nuclear donors to increase their developmental efficiencies and expand their applications in various fields. Peripheral blood cells are useful as ethical and economical cell species because they can be collected easily, even though SCNT embryos derived from hematopoietic cells show poor developmental abilities after reconstruction. Additionally, it is possible to obtain mice that are reactive to specific antigens of interest by using lymphocytes. Although there are still many limitations to the practical use of SCNT, its utilization is steadily expanding.

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Section: Selection Of Appropriate Cell Types For Mouse Scntsupporting

confidence: 62%

Mouse somatic cell nuclear transfer: What has changed and remained unchanged in 25 years

INOUE

2023

Journal of Reproduction and Development

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“…In our previous study, unidirectional freeze‐drying has been shown to impart large axially aligned microchannels in ANAs, thereby improving their dense internal ultrastructure 30,31 . Notably, the freeze‐drying technique itself can be used as a physical decellularization method, effectively lysing cells within tissues and organs 32,33 . Last, nuclease treatments were carried out to ensure adequate clearance of residual antigenic components within ANAs, allowing them to meet the criteria for in vivo transplantation.…”

Section: Introductionmentioning

confidence: 99%

Preparation and assessment of an optimized multichannel acellular nerve allograft for peripheral nerve regeneration

et al. 2022

Bioengineering & Transla Med

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Peripheral nerve regeneration after injury is still a clinical problem. The application of autologous nerve grafting, the gold standard treatment, is greatly restricted. Acellular nerve allografts (ANAs) are considered promising alternatives, but they are difficult to achieve satisfactory therapeutic outcomes, which may be attributed to their compact inherent ultrastructure and substantial loss of extracellular matrix (ECM) components. Regarding these deficiencies, this study developed an optimized multichannel ANA by a modified decellularization method. These innovative ANAs were demonstrated to retain more ECM bioactive molecules and regenerative factors, with effective elimination of cellular antigens. The presence of microchannels with larger pore size allowed ANAs to gain higher porosity and better swelling performance, which improves their internal ultrastructure. Their mechanical properties were more similar to those of native nerves. Moreover, the optimized ANAs exhibited good biocompatibility and possessed significant advantages in supporting the proliferation and migration of Schwann cells in vitro. The in vivo results further confirmed their superior capacity to promote axon regrowth and myelination as well as restore innervation of target muscles, leading to better functional recovery than the conventional ANAs. Overall, this study demonstrates that the optimized multichannel ANAs have great potential for clinical application and offer new insight into the further improvement of ANAs.

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“…Research on the dehydration of somatic cells like lymphocytes or fibroblasts also has helped to make progress in germ cells (Loi et al, 2021; Weng, 2021). The recent birth of healthy cloned offspring derived from freeze‐dried cumulus cells also brings a lot of hope for similar preservation approaches in germ cells (S. Wakayama et al, 2022). Since 2000, sperm cells have been studied in 20 different mammalian species, including humans (Table 1), as ideal candidates for dehydration because of their low water content and compacted DNA.…”

Section: Introductionmentioning

confidence: 99%

Damages and stress responses in sperm cells and other germplasms during dehydration and storage at nonfreezing temperatures for fertility preservation

Comizzoli

Amelkina

Lee

2022

Molecular Reproduction Devel

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Long‐term preservation of sperm, oocytes, and gonadal tissues at ambient temperatures has the potential to lower the costs and simplify biobanking in human reproductive medicine, as well as for the management of animal populations. Over the past decades, different dehydration protocols and long‐term storage solutions at nonfreezing temperatures have been explored, mainly for mammalian sperm cells. Oocytes and gonadal tissues are more challenging to dehydrate so little to no progress have been made. Currently, the detrimental effects of the drying process itself are better characterized than the impact of long‐term storage at nonfreezing temperatures. While structural and functional properties of germ cells can be preserved after dehydration, a long list of damages and stresses in nuclei, organelles, and cytoplasmic membranes have been reported and sometimes mitigated. Characterizing those damages and better understanding the response of germ cells and tissues to the stress of dehydration is fundamental. It will contribute to the development of optimal protocols while proving the safety of alternative storage options for fertility preservation. The objective of this review is to (1) document the types of damages and stress responses, as well as their mitigation in cells dried with different techniques, and (2) propose new research directions.

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Healthy cloned offspring derived from freeze-dried somatic cells

Cited by 14 publications

References 53 publications

Mouse somatic cell nuclear transfer: What has changed and remained unchanged in 25 years

Mouse somatic cell nuclear transfer: What has changed and remained unchanged in 25 years

Preparation and assessment of an optimized multichannel acellular nerve allograft for peripheral nerve regeneration

Damages and stress responses in sperm cells and other germplasms during dehydration and storage at nonfreezing temperatures for fertility preservation

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