Lizard Blastema Organoid Model Recapitulates Regenerated Tail Chondrogenesis

(1) Background: Lizard tail regeneration provides a unique model of blastema-based tissue regeneration for large-scale appendage replacement in amniotes. Green anole lizard ( blastemas contain fibroblastic connective tissue cells (FCTCs), which respond to hedgehog signaling to create cartilage in vi...

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Bibliographic Details
Published in:Journal of developmental biology 2022-02, Vol.10 (1), p.12
Main Authors: Vonk, Ariel C, Hasel-Kolossa, Sarah C, Lopez, Gabriela A, Hudnall, Megan L, Gamble, Darian J, Lozito, Thomas P
Format: Article
Language:English
Subjects:
Amputation
blastema
Cartilage
Cell culture
Chondrogenesis
Collagen
Collagenase
Connective tissue
Connective tissues
Hedgehog protein
hedgehog signaling
Histology
Hybridization
lizard
Mammals
Mimicry
organoid
Organoids
Protocol
regeneration
Tails
Trypsin
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Summary:(1) Background: Lizard tail regeneration provides a unique model of blastema-based tissue regeneration for large-scale appendage replacement in amniotes. Green anole lizard ( blastemas contain fibroblastic connective tissue cells (FCTCs), which respond to hedgehog signaling to create cartilage in vivo. However, an in vitro model of the blastema has not previously been achieved in culture. (2) Methods: By testing two adapted tissue dissociation protocols and two optimized media formulations, lizard tail FCTCs were pelleted in vitro and grown in a micromass blastema organoid culture. Pellets were analyzed by histology and in situ hybridization for FCTC and cartilage markers alongside staged original and regenerating lizard tails. (3) Results: Using an optimized serum-free media and a trypsin- and collagenase II-based dissociation protocol, micromass blastema organoids were formed. Organoid cultures expressed FCTC marker CDH11 and produced cartilage in response to hedgehog signaling in vitro, mimicking in vivo blastema and tail regeneration. (4) Conclusions: Lizard tail blastema regeneration can be modeled in vitro using micromass organoid culture, recapitulating in vivo FCTC marker expression patterns and chondrogenic potential.
ISSN:2221-3759
2221-3759
DOI:10.3390/jdb10010012