Distinct types of fibrocyte can differentiate from mononuclear cells in the presence and absence of serum

49Citations
Citations of this article
63Readers
Mendeley users who have this article in their library.

Abstract

Background: Fibrocytes are bone-marrow derived cells, expressing both haematopoietic and stromal cell markers, which contribute to tissue repair as well as pathological fibrosis. The differentiation of fibrocytes remains poorly characterised and this has limited understanding of their biology and function. In particular two methods are used to generate fibrocytes in vitro that differ fundamentally by the presence or absence of serum. Methodology/Principal Findings: We show here that fibrocytes grown in the absence of serum (SF) differentiate more efficiently from peripheral blood mononuclear cells than CD14+ monocytes, and respond to serum by losing their spindleshaped fibrocyte morphology. Although fibrocytes generated in the presence of serum (SC) express the same range of markers, they differentiate more efficiently from CD14+ monocytes and do not change their morphology in response to serum. Transcriptional analysis revealed that both types of fibrocyte are distinct from each other, fibroblasts and additional monocyte-derived progeny. The gene pathways that differ significantly between SF and SC fibrocytes include those involved in cell migration, immune responses and response to wounding. Conclusions/Significance: These data show that SF and SC fibrocytes are distinct but related cell types, and suggest that they will play different roles during tissue repair and fibrosis where changes in serum proteins may occur. © 2010 Curnow et al.

Figures

  • Figure 1. Differential generation of fibrocytes in serum-free or serum-containing conditions, from PBMC and CD14+ monocytes. Fibrocytes were generated from PBMC (filled circles) or purified CD14+ monocytes (open circles) in either serum-free (SF) or serum-containing (SC; 20% HIFCS). The number of fibrocytes is shown during an 18-day culture period (A,B; mean 6 sd of triplicate culture wells). Error bars are only shown in one direction for clarity. Bar represents 50 mm. Fibrocytes and macrophages (MØ) were generated from a number of different normal healthy volunteers (minimum of n = 6) and the number of cells per well counted at day 11 of culture (C). * = p,0.05; *** = p,0.001; Mann-Whitney test. doi:10.1371/journal.pone.0009730.g001
  • Figure 2. Both serum-free and serum-containing fibrocytes express both haematopoietic and stromal cell markers. Fibrocytes were generated from PBMC under serum-free (SF) and serum-containing (SC) culture conditions, as well as macrophages. Fibroblast cell lines were included as a positive control for stromal cell markers. Immunostaining is shown in red for the stromal markers fibronectin, collagens I and III and
  • Figure 3. Addition of serum to serum-free generated fibrocytes results in a loss of fibrocyte morphology. Fibrocytes generated from PBMC under serum-free (SF) culture conditions were cultured in the presence of serum-containing (SC) culture medium, and video recordings taken for a period of 150 min. Still photographs are shown after each 30 min period (A), with arrows indicating individual cells that have lost the typical elongated fibrocyte morphology, taking on a more rounded appearance. Bar represents 50 mm. The serum-free culture conditions were changed to serum-containing conditions after 4, 8, 11 and 14 days as indicated by the dotted line (B), with a reciprocal experiment where serum-containing medium was changed to serum-free culture conditions (C). Data are the mean 6 sd of triplicate culture wells and are representative of three separate experiments. The median number of cells with fibrocyte morphology at 24 h after the change of culture conditions from multiple experiments is shown (D). *** = p,0.001; NS = not significant, p.0.05; Mann-Whitney test. Error bars are only shown in one direction for clarity. doi:10.1371/journal.pone.0009730.g003
  • Figure 4. Fibrocytes generated in serum-free and serum-containing conditions show distinct transcriptional profiles from each other, as well as other monocyte-derived progeny. The expression of 35,788 genes was determined by gene microarray analysis for fibroblasts,
  • Table 1. Pathways identified by DAVID analysis of the two-way SAM gene list of serum-free versus serum-containing fibrocytes.

References Powered by Scopus

Get full text
Get full text

Cited by Powered by Scopus

Get full text

This article is free to access.

This article is free to access.

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Cite

CITATION STYLE

APA

Curnow, S. J., Fairclough, M., Schmutz, C., Kissane, S., Denniston, A. K. O., Nash, K., … Salmon, M. (2010). Distinct types of fibrocyte can differentiate from mononuclear cells in the presence and absence of serum. PLoS ONE, 5(3). https://doi.org/10.1371/journal.pone.0009730

Readers over time

‘10‘11‘12‘13‘14‘15‘16‘17‘18‘19‘20‘21‘22‘23‘24036912

Readers' Seniority

Tooltip

PhD / Post grad / Masters / Doc 27

59%

Researcher 15

33%

Professor / Associate Prof. 3

7%

Lecturer / Post doc 1

2%

Readers' Discipline

Tooltip

Medicine and Dentistry 22

45%

Agricultural and Biological Sciences 17

35%

Biochemistry, Genetics and Molecular Bi... 6

12%

Immunology and Microbiology 4

8%

Save time finding and organizing research with Mendeley

Sign up for free
0