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001-es BibID:	BIBFORM113275
035-os BibID:	(cikkazonosító)e835 (Scopus)85164146982
Első szerző:	Takács Roland Ádám (molekuláris biológus, biokémikus)
Cím:	Isolation and Micromass Culturing of Primary Chicken Chondroprogenitor Cells for Cartilage Regeneration / Takács Roland, Juhász Tamás, Katona Éva, Somogyi Csilla, Vágó Judit, Hajdú Tibor, Barna Krisztina Bíróné, Nagy Péter, Zákány Róza, Matta Csaba
Dátum:	2023
ISSN:	2691-1299
Megjegyzések:	Much of the skeletal system develops by endochondral ossification, a process that takes place in early fetal life. This makes the early stages of chondrogenesis, i.e., when chondroprogenitor mesenchymal cells differentiate to chondroblasts, challenging to study in vivo. In vitro methods for the study of chondrogenic differentiation have been available for some time. There is currently high interest in developing fine-tuned methodology that would allow chondrogenic cells to rebuild articular cartilage and restore joint functionality. The micromass culture system that relies on embryonic limb bud-derived chondroprogenitor cells is a popular method for the study of the signaling pathways that control the formation and maturation of cartilage. In this protocol, we describe a technique fine-tuned in our laboratory for culturing limb bud-derived mesenchymal cells from early-stage chick embryos in high density (Basic Protocol 1). We also provide a fine-tuned method for high-efficiency transient transfection of cells before plating using electroporation (Basic Protocol 2). In addition, protocols for histochemical detection of cartilage extracellular matrix using dimethyl methylene blue, Alcian blue, and safranin O are also provided (Basic Protocol 3 and Alternate Protocols 1 and 2, respectively). Finally, a step-by-step guide on a cell viability/proliferation assay using MTT reagent is also described (Basic Protocol 4). © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Micromass culture of chick embryonic limb bud-derived cells. Basic Protocol 2: Transfection of cells with siRNA constructs using electroporation prior to micromass culturing. Basic Protocol 3: Qualitative and quantitative assessment of cartilage matrix production using dimethyl methylene blue staining and image analysis. Alternate Protocol 1: Qualitative assessment of cartilage matrix production using Alcian blue staining. Alternate Protocol 2: Qualitative assessment of cartilage matrix production using safranin O staining. Basic Protocol 4: Measurement of mitochondrial activity with the MTT assay.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban folyóiratcikk
Megjelenés:	Current Protocols. - 3 : 7 (2023), p. 1-29. -
További szerzők:	Juhász Tamás (1976-) (biológus, orvosbiológus) Katona Éva (1986-) (molekuláris biológus) Somogyi Csilla (1983-) (biológus, angol-magyar szakfordító) Vágó Judit (1990-) (molekuláris biológus) Hajdú Tibor (1988-) (általános orvos) Barna Krisztina B. Nagy Péter (1971-) (biofizikus) Zákány Róza (1963-) (anatómus-, kötőszövetbiológus) Matta Csaba (1980-) (molekuláris biológus, genetikus, angol szakfordító)
Pályázati támogatás:	OTKA-134304 OTKA COST Egyéb
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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