Macrophage Colony Stimulating Factor-Induced Macrophage Differentiation Influences Myotube Elongation

Abstract

Background:

Unaccustomed exercise, high-intensity dynamic sports activities, or the resumption of normal weight-bearing after a period of disuse can induce skeletal muscle injury, which activates an inflammatory response followed by muscle regeneration. Specific subsets of macrophages are involved in muscle regeneration. But the exact role of macrophage differentiation during muscle regeneration remains to be elucidated.

Objective:

The objective of the study was to examine the effect of macrophage colony stimulating factor (M-CSF)-differentiated, lipopolysaccharides (LPS)-stimulated-macrophage-conditioned medium on muscle-cell proliferation, fusion, and elongation, which are key events during muscle regeneration and myogenesis.

Method:

Murine C2C12 myoblasts were cultured in conditioned medium obtained from PU5-1R macrophages that were (a) undifferentiated, unstimulated; (b) M-CSF-differentiated, unstimulated; (c) undifferentiated, LPS-stimulated; or (d) M-CSF-differentiated, LPS-stimulated. Myoblast proliferation ratio, nuclei number, and length were measured.

Results:

C2C12 cells cultured in conditioned medium from M-CSF-differentiated, LPS-stimulated macrophages had significantly more nuclei and greater length than cells cultured in conditioned medium from undifferentiated, LPS-stimulated macrophages. Dilution and denaturization of the M-CSF-differentiated, LPS-stimulated-macrophage medium prevented a marked increase in C2C12 nuclei number and length. However, the C2C12 myoblast proliferation ratio was significantly greater in conditioned medium from undifferentiated, LPS-stimulated macrophages than in conditioned medium from M-CSF-differentiated, LPS-stimulated macrophages.

Conclusions:

M-CSF-differentiated, LPS-stimulated macrophages may influence myogenesis and the early and terminal stages of muscle regeneration. This knowledge may aid in developing therapies that will directly expedite muscle repair and lead to faster rehabilitation and reduced rehabilitation costs.

Keywords

C2C12 myoblasts PU5-1R macrophages colony stimulating factor myogenesis myotube fusion

Get full access to this article

View all access options for this article.

References

Arkins

Rebeiz

Biragyn

Reese

D. L.

Kelley

K. W.

(1993). Murine macrophages express abundant insulin-like growth factor-I class I Ea and Eb transcripts. Endocrinology, 133, 2334–2343.

Arnold

Henry

Poron

Baba-Amer

Van Rooijen

Plonquet

A.,

…Chazaus

(2007). Inflammatory monocytes recruited after skeletal muscle injury switch into antiinflammatory macrophages to support myogenesis. Journal of Experimental Medicine, 204, 1057–1069.

Becker

Warren

M. K.

Haskill

(1987). Colony-stimulating factor-induced monocyte survival and differentiation into macrophages in serum-free cultures. Journal of Immunology, 139, 3703–3709.

Borycki

A. G.

Smadja

Stanley

Leibovitch

S. A.

(1995). Colony-stimulating factor 1 (CSF-1) is involved in an autocrine growth control of rat myogenic cells. Experimental Cell Research, 218, 213–222.

Brugger

Kreutz

Andreesen

(1991). Macrophage colony-stimulating factor is required for human monocyte survival and acts as a cofactor for their terminal differentiation to macrophages in vitro. Journal of Leukocyte Biology, 49, 483–488.

Cantini

Giurisato

Radu

Tiozzo

Pampinella

Senigaglia

D.,

… Vitiello

(2002). Macrophage-secreted myogenic factors: A promising tool for greatly enhancing the proliferative capacity of myoblasts in vitro and in vivo. Neurological Sciences, 23, 189–194.

Cantini

Massimino

M. L.

Bruson

Catani

Dalla Libera

Carraro

(1994). Macrophages regulate proliferation and differentiation of satellite cells. Biochemical and Biophysical Research Communications, 202, 1688–1696.

Cheng

Nguyen

M. H.

Fantuzzi

Koh

T. J.

(2008). Endogenous interferon-γ is required for efficient skeletal muscle regeneration. American Journal of Physiology—Cell Physiology, 294, C1183–C1191.

Chitu

Stanley

E. R.

(2006). Colony-stimulating factor-1 in immunity and inflammation. Current Opinion in Immunology, 18, 39–48.

10.

Darwich

Coma

Pena

Bellido

Blanco

E. J. J.

Este

J. A.

… Bofill

(2009). Secretion of interferon-γ by human macrophages demonstrated at the single-cell level after costimulation with interleukin (IL)-12 plus IL-18. Immunology, 126, 386–393.

11.

Evans

Kamdar

S. J.

Fuller

J. A.

Krupke

D. M.

(1995). The potential role of the macrophage colony-stimulating factor, CSF-1, in inflammatory responses: Characterization of macrophage cytokine gene expression. Journal of Leukocyte Biology, 58, 99–107.

12.

Ferri

Barbieri

Burattini

Guescini

D'Emilio

Biagiotti

… Falcieri

(2009). Expression and subcellular localization of myogenic regulatory factors during the differentiation of skeletal muscle C2C12 myoblasts. Journal of Cellular Biochemistry, 108, 1302–1317.

13.

Fleetwood

A. J.

Lawrence

Hamilton

J. A.

Cook

A. D.

(2007). Granulocyte-macrophage colony stimulating factor (CSF) and macrophage CSF-dependent macrophage phenotypes display differences in cytokine profiles and transcription factor activities: Implications for CSF blockade in inflammation. Journal of Immunology, 178, 5245–5252.

14.

Hanamura

Asakura

Tanabe

(1997). Macrophage colony-stimulating factor (M-CSF) augments cytokine induction by lipopolysaccharide (LPS)-stimulation and by bacterial infections in mice. Immunopharmacology, 37, 15–23.

15.

Kamdar

S. J.

Chapoval

A. I.

Phelps

Fuller

J. A.

Evans

(1996). Differential sensitivity of mouse mononuclear phagocytes to CSF-1 and LPS: The potential in vivo relevance of enhanced IL-6 gene expression. Cell Immunology, 174, 165–172.

16.

Kamdar

S. J.

Fuller

J. A.

Nishikawa

S. I.

Evans

(1997). Priming of mouse macrophages with the macrophage colony-stimulating factor (CSF-1) induces a variety of pathways that regulate expression of the interleukin 6 (Il6) and granulocyte-macrophage colony-stimulating factor (Csfgm) genes. Experimental Cell Research, 235, 108–116.

17.

Koh

T. J.

Bryer

S. C.

Pucci

A. M.

Sisson

T. H.

(2005). Mice deficient in plasminogen activator inhibitor-1 have improved skeletal muscle regeneration. American Journal of Physiology—Cell Physiology, 289, C217–223.

18.

Leenen

P. J. M.

de Bruijn

M. F. T. R.

Voerman

J. S. A.

Campbell

P. A.

van Ewijk

(1994). Markers of mouse macrophage development detected by monoclonal antibodies. Journal of Immunological Methods, 174, 5–19.

19.

Leenen

P. J. M.

Melis

Kraal

Hoogeveen

A. T.

van Ewijk

(1992). The monoclonal antibody ER-BMDM1 recognizes a macrophage and dendritic cell differentiation antigen with aminopeptidase activity. European Journal of Immunology, 22, 1567–1572.

20.

Leibovitch

S. A.

Leibovitch

M. P.

Borycki

A. G.

Harel

(1989). Expression of CSF-1-receptor-related proteins in muscular stem cells. Oncogene Research, 4, 157–162.

21.

Massimino

M. L.

Rapizzi

Cantini

Libera

L. D.

Mazzoleni

Arslan

Carraro

(1997). ED2+ macrophages increase selectively myoblast proliferation in muscle cultures. Biochemical and Biophysical Research Communications, 235, 754–759.

22.

Riss

Moravec

(1996). Improved non-radioactive assay to measure cellular proliferation or toxicity: The CellTiter 96^® AQ_ueous One Solution Cell Proliferation Assay. Promega Notes Magazine, 59, 19–23.

23.

Stout

R. D.

Jiang

Matta

Tietzel

Watkins

S. K.

Suttles

(2005). Macrophages sequentially change their functional phenotype in response to changes in microenvironmental influences. Journal of Immunology, 175, 342–349.

24.

Pierre

B. A.

St. Tidball

J. G.

(1994). Differential response of macrophage subpopulations to soleus muscle reloading after rat hindlimb suspension. Journal of Applied Physiology, 77, 290–297.

25.

Pierre

Schneider B.

St. Correia

L. A.

Cannon

J. G.

(1999). Sex differences in leukocyte invasion in injured murine skeletal muscle. Research in Nursing & Health, 22, 243–250.

26.

Sweet

M. J.

Campbell

C. C.

Sester

D. P.

McDonald

R. C.

Stacey

K. J.

… Liew

F. Y.

(2002). Colony-stimulating factor-1 suppresses responses to CpG DNA and expression of Toll-like receptor 9 but enhances responses to lipopolysaccharide in murine macrophages. Journal of Immunology, 168, 392–399.

27.

Sweet

M. J.

Hume

D. A.

(2003). CSF-1 as a regulator of macrophage activation and immune responses. Archivum Immunologiae et Therapiae Experimentalis, 51, 169–177.

28.

Tidball

J. G.

Villalta

S. A.

(2010). Regulatory interactions between muscle and the immune system during muscle regeneration. American Journal of Physiology—Regulatory Integrative and Comparative Physiology, 298, R1173–R1187.

29.

Tidball

J. G.

Wehling-Henricks

(2007). Macrophages promote muscle membrane repair and muscle fibre growth and regeneration during modified muscle loading in mice. in vivo. Journal of Physiology, 578, 327–336.

30.

Valovich McLeod

T. C.

Bay

R. C.

Parsons

J. T.

Sauers

E. L.

Snyder

A. R.

(2009). Recent injury and health-related quality of life in adolescent athletes. Journal of Athletic Training, 44, 603–610.

31.

Verreck

F. A. W.

de boer

Langenberg

D. M. L.

van der Zanden

Ottenhoff

T. H. M.

(2006). Phenotypic and functional profiling of human proinflammatory type-1 and anti-inflammatory type-2 macrophages in response to microbial antigens and IFN-γ- and CD40L-mediated costimulation. Journal of Leukocyte Biology, 79, 285–293.