Dataset Information

The proteome of osteoblasts in a 3D culture bioreactor model with static and perfusion conditions using biomimicry

ABSTRACT: Disorders influencing bone form a major burden worldwide, with osteoporosis being the most common fracture-inducing bone disease. Currently, animal models are being utilized to develop and screen novel treatments. However, due to interspecies variations, different microenvironments, and ethical considerations, there is a need for a complex 3D bone model. This research aimed to perform a pilot study on developing a standardized representative bone model using a perfusion-based bioreactor, and to characterize the overall 3D bone proteomic profile. An immortalized human fetal osteoblastic cell line (hFOB1.19) was seeded on collagen scaffolds and cultured for 21 days in a perfusion bioreactor system and in static cultures. Different analyses allowed the monitoring of cell viability and compared morphological and proteome differences between both conditions. Scanning electron microscopy images suggested an altered cellular morphology within the bioreactor culture compared to the static cultures. A proteomic analysis identified a total of 1897 proteins and a total of 93 unique regulated proteins. Enrichment analyses of these proteins revealed seven significant pathways including TNF-alpha signaling pathway via NF-kB. These pathways also included hypoxia and epithelial-mesenchymal transition, where the EMT pathway was the most significant and indicated a reduction in cell motility within the bioreactor system on day 21. This preliminary study suggests that the U-CUP perfusion bioreactor is promising for further exploration of facilitating osteogenic differentiation induction in 3D cultures.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Akos Vegvari

LAB HEAD: Akos Vegvari

PROVIDER: PXD057937 | Pride | 2025-05-07

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20230225_HF2_01_KaiB_B6.raw	Raw
	20230225_HF2_02_KaiB_B5.raw	Raw
	20230225_HF2_03_KaiB_B8.raw	Raw
	20230225_HF2_04_KaiB_B7.raw	Raw
	20230225_HF2_05_KaiB_S1.raw	Raw

Items per page:

1 - 5 of 9

Publications

The proteome of osteoblasts in a 3D culture perfusion bioreactor model compared with static conditions.

Radi Sonya S EzEldeen Mostafa M Végvári Ákos Á Coates Dawn D Jacobs Reinhilde R Bostanci Nagihan N Bao Kai K

Scientific reports 20250409 1

Bone disorders represent a significant global burden. Currently, animal models are used to develop and screen novel treatments. However, interspecies variations and ethical concerns highlight the need for a more complex 3D bone model. In this study, we developed a simplified in vitro bone-like model using a U-CUP perfusion-based bioreactor system, designed to provide continuous nutrient flow and mechanostimulation through 3D cultures. An immortalized human fetal osteoblastic cell line was seeded ...[more]

PMID: 40204872

Similar Datasets

Project description:Development of systems allowing the maintenance of native properties of mesenchymal stromal cells (MSC) is a critical challenge for studying physiological functions of skeletal progenitors, as well as towards cellular therapy and regenerative medicine applications. Conventional stem cell culture in monolayer on plastic dishes (2D) is associated with progressive loss of functionality, likely due to the absence of a biomimetic microenvironment and the selection of adherent populations. Here we demonstrate that 2D MSC expansion can be entirely bypassed by culturing freshly isolated bone marrow cells within the pores of 3D scaffolds in a perfusion-based bioreactor system, followed by enzymatic digestion for cell retrieval. The 3D-perfusion system supported MSC growth while maintaining cells of the hematopoietic lineage, and thus generated a cellular environment mimicking some features of the bone marrow stroma. As compared to 2D-expansion, sorted CD45- cells derived from 3D-perfusion culture after the same time (3 weeks) or a similar extent of proliferation (7-8 doublings) maintained a 4.3-fold higher clonogenicity and exhibited a superior differentiation capacity towards all typical mesenchymal lineages, with similar immunomodulatory function in vitro. Transcriptomic analysis performed on MSC from 5 donors validated the robustness of the process and indicated a reduced inter-donor variability as well as a significant upregulation of multipotency-related gene clusters following 3D-perfusion as compared to 2D expansion. The described system offers a model to study how factors of a 3D engineered niche may regulate MSC function and, by streamlining conventional labor-intensive processes, is prone to automation and scalability within closed bioreactor systems. Nucleated cells were isolated from 5 fresh human bone marrow aspirates by means of red blood cells lyses buffer and then were seeded into a 3D perfusion bioreactor system using a pure hydroxyapatite 3D scaffold and in conventional Petri dishes (2D). After culture for 19 days, cells from both systems were enzymatically retrieved and sorted using anti-CD45-coated magnetic beads. Total RNA was extracted from CD45- cells, QCed and hybridized to Affymetrix microarrays.