Optimization of the static human osteoblast/osteoclast co-culture system

James Jam Jolly, Kok Yong Chin, Mohd Fozi Nur Farhana, Ekram Alias, Kien Hui Chua, Wan Nuraini Wan Hasan, Soelaiman Ima-Nirwana

Research output: Contribution to journalArticle

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Abstract

Osteoblasts (OBs) and osteoclasts (OCs) are 2 major groups of bone cells. Their cell-to-cell interactions are important to ensure the continuity of the bone-remodeling process. Therefore, the present study was carried out to optimize an OB/OC co-culture system utilizing the human OB cell line hFOB 1.19 and OCs extracted from peripheral blood mononuclear cells (PBMNCs). It was a 2-step procedure, involving the optimization of the OB culture and the co-culture of the OBs with PBMNCs at an optimum ratio. Firstly, pre-OBs were cultured to 90% confluency and the time required for differentiation was determined. OB differentiation was determined using the van Gieson staining to detect the presence of collagen and Alizarin Red for calcium. Secondly, OBs and OCs were co-cultured at the ratios of 1 OC: 1 OB, 1 OC: 4 OBs, 2 OCs: 1 OB, and 1 OC: 2 OBs. Tartrate-resistant acid phosphatase (TRAP) staining was used to detect the differentiation of the OCs. The results showed that collagen was present on day 1, whereas calcium was detected as early as day 3. Based on the result of TRAP staining, 1 OC: 2 OBs was taken as the most appropriate ratio. No macrophage colony-stimulating factor and receptor activator of the nuclear factor-κB ligand were added because they were provided by the OBs. In conclusion, these optimization processes are vital as they ensure the exact time point and ratio of the OB/OC co-culture in order to produce a reliable and reproducible co-culture system.

LanguageEnglish
Pages208-213
Number of pages6
JournalIranian Journal of Medical Sciences
Volume43
Issue number2
Publication statusPublished - 1 Mar 2018

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Osteoclasts
Coculture Techniques
Osteoblasts
Staining and Labeling
Blood Cells
Collagen
Macrophage Colony-Stimulating Factor Receptors
Calcium
Bone Remodeling
Cell Communication

Keywords

  • Bone and Bones
  • Bone Remodeling
  • Coculture Techniques
  • Osteoblasts
  • Osteoclasts

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Optimization of the static human osteoblast/osteoclast co-culture system. / Jolly, James Jam; Chin, Kok Yong; Farhana, Mohd Fozi Nur; Alias, Ekram; Chua, Kien Hui; Hasan, Wan Nuraini Wan; Ima-Nirwana, Soelaiman.

In: Iranian Journal of Medical Sciences, Vol. 43, No. 2, 01.03.2018, p. 208-213.

Research output: Contribution to journalArticle

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