Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab

Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab

Abstract Introduction and objectives: osteoclasts are terminally differentiated giant multinucleated cells derived from the fusion of mononuclear progenitors of the monocyte/macrophage hematopoietic lineage. The objective of our group was to achieve the best method for osteoclast differentiation, from both RAW 264.7 cells and peripheral blood monocytes. Material and methods: RAW 264.7 cells and human PBMCs were differentiated into osteoclasts. Success in differentiation was assessed by TRAP staining. Osteoclast activity was detected by the resorption pits in Corning® Osteo Assay Surface Plates. Results: the optimal cell density for RAW 264.7 cell differentiation was 25,000 cells/cm2 with 30 ng/mL of RANKL for 6 days. Osteoclasts differentiated from PBMCs were observed after 4 weeks with 25 ng/mL M-CSF and 30 ng/mL RANKL. Individual pits or multiple pit clusters were observed on the surface plates. Conclusions: we report optimal conditions for the differentiation of osteoclasts from

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Main Authors: Jurado,Susana, Parés,Albert, Peris,Pilar, Combalia,Andrés, Monegal,Ana, Guañabens,Nuria
Format: Digital revista
Language:English
Published: Sociedad Española de Investigaciones Óseas y Metabolismo Mineral 2023
Online Access:https://scielo.isciii.es/scielo.php?script=sci_arttext&pid=S1889-836X2023000100002
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spelling oai:scielo:S1889-836X20230001000022023-05-25Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our labJurado,SusanaParés,AlbertPeris,PilarCombalia,AndrésMonegal,AnaGuañabens,Nuria Osteoclasts Differentiation Bone resorption Abstract Introduction and objectives: osteoclasts are terminally differentiated giant multinucleated cells derived from the fusion of mononuclear progenitors of the monocyte/macrophage hematopoietic lineage. The objective of our group was to achieve the best method for osteoclast differentiation, from both RAW 264.7 cells and peripheral blood monocytes. Material and methods: RAW 264.7 cells and human PBMCs were differentiated into osteoclasts. Success in differentiation was assessed by TRAP staining. Osteoclast activity was detected by the resorption pits in Corning® Osteo Assay Surface Plates. Results: the optimal cell density for RAW 264.7 cell differentiation was 25,000 cells/cm2 with 30 ng/mL of RANKL for 6 days. Osteoclasts differentiated from PBMCs were observed after 4 weeks with 25 ng/mL M-CSF and 30 ng/mL RANKL. Individual pits or multiple pit clusters were observed on the surface plates. Conclusions: we report optimal conditions for the differentiation of osteoclasts fromSociedad Española de Investigaciones Óseas y Metabolismo MineralRevista de Osteoporosis y Metabolismo Mineral v.15 n.1 20232023-03-01journal articletext/htmlhttps://scielo.isciii.es/scielo.php?script=sci_arttext&pid=S1889-836X2023000100002en
institution SCIELO
collection OJS
country España
countrycode ES
component Revista
access En linea
databasecode rev-scielo-es
tag revista
region Europa del Sur
libraryname SciELO
language English
format Digital
author Jurado,Susana
Parés,Albert
Peris,Pilar
Combalia,Andrés
Monegal,Ana
Guañabens,Nuria
spellingShingle Jurado,Susana
Parés,Albert
Peris,Pilar
Combalia,Andrés
Monegal,Ana
Guañabens,Nuria
Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab
author_facet Jurado,Susana
Parés,Albert
Peris,Pilar
Combalia,Andrés
Monegal,Ana
Guañabens,Nuria
author_sort Jurado,Susana
title Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab
title_short Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab
title_full Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab
title_fullStr Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab
title_full_unstemmed Osteoclast generation from RAW 264.7 and PBMC cells. The set up in our lab
title_sort osteoclast generation from raw 264.7 and pbmc cells. the set up in our lab
description Abstract Introduction and objectives: osteoclasts are terminally differentiated giant multinucleated cells derived from the fusion of mononuclear progenitors of the monocyte/macrophage hematopoietic lineage. The objective of our group was to achieve the best method for osteoclast differentiation, from both RAW 264.7 cells and peripheral blood monocytes. Material and methods: RAW 264.7 cells and human PBMCs were differentiated into osteoclasts. Success in differentiation was assessed by TRAP staining. Osteoclast activity was detected by the resorption pits in Corning® Osteo Assay Surface Plates. Results: the optimal cell density for RAW 264.7 cell differentiation was 25,000 cells/cm2 with 30 ng/mL of RANKL for 6 days. Osteoclasts differentiated from PBMCs were observed after 4 weeks with 25 ng/mL M-CSF and 30 ng/mL RANKL. Individual pits or multiple pit clusters were observed on the surface plates. Conclusions: we report optimal conditions for the differentiation of osteoclasts from
publisher Sociedad Española de Investigaciones Óseas y Metabolismo Mineral
publishDate 2023
url https://scielo.isciii.es/scielo.php?script=sci_arttext&pid=S1889-836X2023000100002
work_keys_str_mv AT juradosusana osteoclastgenerationfromraw2647andpbmccellsthesetupinourlab
AT paresalbert osteoclastgenerationfromraw2647andpbmccellsthesetupinourlab
AT perispilar osteoclastgenerationfromraw2647andpbmccellsthesetupinourlab
AT combaliaandres osteoclastgenerationfromraw2647andpbmccellsthesetupinourlab
AT monegalana osteoclastgenerationfromraw2647andpbmccellsthesetupinourlab
AT guanabensnuria osteoclastgenerationfromraw2647andpbmccellsthesetupinourlab
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