Epigenetic drug 5-azacytidine impairs the potential for odontogenesis but improves tooth morphogenesis in the transplanted embryonic mandible

Radujković, Vedran; Himelreich-Perić, Marta; Takahashi, Marta; Sinčić, Nino; Bojana, Ana Katušić; Ježek, Davor; Jurić-Lekić, Gordana; Bulić-Jakuš, Floriana

doi:10.18054/pb.v121-122i1-2.10718

Cited by 2 publications

(1 citation statement)

References 16 publications

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“…In contrast to some experiments with dissociated cells, where necessary hydrogel encapsulation heavily influenced bone‐forming potential in the subcutaneous ectopic assay in vivo (Verbeeck et al, 2019), we used natural encapsulation by the kidney capsule. The latter technique has been successfully used before for direct transplantation of parts of rodent embryos, epiblast, three germ‐layer gastrulating embryos‐proper, epiglottis, and mandible which always continued their development in such a secluded and well‐vascularized space (Bulic‐Jakus et al, 2016; Marinovic‐Kulisic et al, 2011; Radujkovic et al, 2020). It was described recently that subcapsular kidney space was not optimal for differentiation of cartilage engineered from human embryonic stem cells in specific chemically defined serum‐free conditions because it gave rise also to the bone in contrast to the subcutaneous transplantation (Yang et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Transmembranous and enchondral osteogenesis in transplants of rat limb buds cultivated in serum‐ and protein‐free culture medium

Himelreich-Perić¹,

Mužić-Radović²,

Marić³

et al. 2022

Anat Histol Embryol

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Cartilage differentiates in rat limb buds cultivated in a chemically defined protein‐free medium in the same manner as in the richer serum‐supplemented medium. We aimed to investigate the remaining differentiation potential of pre‐cultivated limb buds by subsequent transplantation in vivo. Rat front (FLBs) and hind‐limb buds (HLBs) were isolated from Fischer rat dams at the 14th gestation day (GD 14) and cultivated at the air‐liquid interface in Eagle's Minimum Essential Medium (MEM) alone; with 5 μM of 5‐azacytidine (5azaC) or with rat serum (1:1). Overall growth was measured seven times during the culture by an ocular micrometre. After 14 days, explants were transplanted under the kidney capsule of adult males. Growth of limb buds was significantly lower in all limb buds cultivated in MEM than in those cultivated with serum. In MEM with 5azaC, growth of LBs was significantly lower only on day 3 of culture. Afterwards, it was higher throughout the culture period, although a statistically significant difference was assessed only for HLBs. In transplants, mixed structures developed with the differentiated transmembranous bone, cartilage with enchondral ossification, bone‐marrow, sebaceous gland, and hair that have never been found in vitro. Nerves differentiated only in transplants precultivated in the serum‐supplemented medium. We conclude that pre‐cultivation of LBs in a chemically defined protein‐free medium does not restrict osteogenesis and formation of epidermal appendages but is restrictive for neural tissue. These results are important for understanding limb development and regenerative medicine strategies.

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