Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Jun 6;69(3):129-138.
doi: 10.1262/jrd.2022-105. Epub 2023 Mar 16.

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

Kimiko Inoue  1   2
Affiliations

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

Kimiko Inoue. J Reprod Dev. .

Abstract

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.

Keywords: Genome reprogramming; Imprinted gene; Mouse; Peripheral blood cell; Somatic cell nuclear transfer.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest associated with this manuscript.

Figures

Fig. 1.
Fig. 1.
High developmental abilities of SCNT embryos derived from somatic cells in 129 inbred or hybrid strains. (A) Twelve newborn pups cloned from (DBA/2 × 129/Sv-ter) F1 immature Sertoli cells. (B) The birth rates of SCNT embryos in 129 inbred or hybrid strains. 129×B6: (129/Sv-ter × C57BL/6) F1, B6×129: (C57BL/6 × 129/Sv-ter) F1, 129: 129/Sv-ter, 129×129: (129/Sv-ter × 129/SvJ) F1, DBA × 129: (DBA/2 × 129/Sv-ter) F1. The figures were modified from a previous report [28].
Fig. 2.
Fig. 2.
SCNT fetuses derived from various blood cells. (A) SCNT fetuses from NKT cells. Two fetuses grew into fertile adults. (B) SCNT fetuses from LT-HSC cells collected from (B6×129/Sv-ter) F1 mouse strain. (C) SCNT fetuses from T-cell lymphocytes reactive to a specific antigen (D. farinae). (D) Histopathological changes in the lungs of mice possessing rearranged TCRα and/or TCRβ after a few D. farinae antigen challenges. H&E: hematoxylin and eosin stain; PAS: periodic acid-Schiff. Scale bar: 50 μm. The figures were modified from previous reports [47, 56, 59].
Fig. 3.
Fig. 3.
Aberrant Xist gene expression in SCNT embryos. (A) Ectopic Xist expression in blastomeres of SCNT embryos. In IVF embryos, one or no Xist signals were detected in female and male embryos, respectively. In SCNT embryos, two or single signals by ectopic expression from active X chromosomes were observed in female and male embryos, respectively. (B) Global gene expression on the X chromosome of IVF and cumulus SCNT embryos. Green: IVF blastocysts; Red: SCNT blastocysts. Expression levels were repressed in SCNT embryos. (C) SCNT fetuses cloned from WT (left) and Xist KO donor cells (right). The figures were modified from a previous report [73].
Fig. 4.
Fig. 4.
Normalization of SCNT placental hyperplasia by ameliorating C2MC expression levels. C2MC was overexpressed in SCNT placentas due to LOI expression of both alleles. SCNT placental hyperplasia was normalized in mice cloned from C2MC maternal allele-deleted donor cells by ameliorating C2MC expression. The figures have been modified from a previous report [82].
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Briggs R, King TJ. Transplantation of living nuclei from blastula cells into enucleated frogs’ eggs. Proc Natl Acad Sci USA 1952; 38: 455–463. - PMC - PubMed
    1. Gurdon JB, Elsdale TR, Fischberg M. Sexually mature individuals of Xenopus laevis from the transplantation of single somatic nuclei. Nature 1958; 182: 64–65. - PubMed
    1. Gurdon JB. Adult frogs derived from the nuclei of single somatic cells. Dev Biol 1962; 4: 256–273. - PubMed
    1. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KH. Viable offspring derived from fetal and adult mammalian cells. Nature 1997; 385: 810–813. - PubMed
    1. Wakayama T, Perry ACF, Zuccotti M, Johnson KR, Yanagimachi R. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 1998; 394: 369–374. - PubMed
  NODES
Idea 1
idea 1
twitter 2