Biomedical Science and Engineering
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Biomedical Science and Engineering. 2014, 2(1), 5-12
DOI: 10.12691/bse-2-1-2
Open AccessReview Article

Different in Vitro Activation Methods for Latent Transforming Growth Factors (TGF)–β: Considerable Exogenous Factors to Promote Higher Mesenchymal-Origin Cell Proliferation in a Bioprocessing Platform

Partha S. Saha1, and Michael Doran2

1Department of Biomedical Sciences, Faculty of Medicine, University of Leuven (KU Leuven), Leuven, Belgium

2School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT), Brisbane, Australia

Pub. Date: February 12, 2014

Cite this paper:
Partha S. Saha and Michael Doran. Different in Vitro Activation Methods for Latent Transforming Growth Factors (TGF)–β: Considerable Exogenous Factors to Promote Higher Mesenchymal-Origin Cell Proliferation in a Bioprocessing Platform. Biomedical Science and Engineering. 2014; 2(1):5-12. doi: 10.12691/bse-2-1-2


Regenerative medicine includes two efficient techniques, namely tissue-engineering and cell-based therapy in order to repair tissue damage efficiently. Most importantly, huge numbers of autologous cells are required to deal these practices. Nevertheless, primary cells, from autologous tissue, grow very slowly while culturing in vitro; moreover, they lose their natural characteristics over prolonged culturing period. Transforming growth factors-beta (TGF-β) is a ubiquitous protein found biologically in its latent form, which prevents it from eliciting a response until conversion to its active form. In active form, TGF-β acts as a proliferative agent in many cell lines of mesenchymal origin in vitro. This article reviews on some of the important activation methods-physiochemical, enzyme-mediated, non-specific protein interaction mediated, and drug-induced- of TGF-β, which may be established as exogenous factors to be used in culturing medium to obtain extensive proliferation of primary cells.

latent TGF-β activation extensive cell culture exogenous factors hMSC proliferation bioreactor

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[1]  Serra M, Brito C and Alves PM. “Bioengineering strategies for stem cell expansion and differentiation.” Canal Bioquímica; 7: 30‐38. 2010.
[2]  Castro-Malaspina H, Gay RE, Resnick G, Kapoor N, Meyers P, Chiarieri D, McKenzie S, Broxmeyer HE, Moore MA. “Characterization of human bone marrow fibroblast colony-forming cells (CFU-F) and their progeny.” Blood; 56 (2): 289-301. 1980.
[3]  Fehrer C and Lepperdinger G. “Mesenchymal stem cell aging.” Experimental Gerontology; 40: 926-930. 2005.
[4]  Todaro GJ. “Autocrine secretion of peptide growth factors by tumor cells.” Natl. Cancer Inst. Monogr; 60: 139-147. 1982.
[5]  Lyons RM, Keski-Oja J, and Moses HL. “Proteolytic Activation of Latent Transforming Growth Factor-β from Fibroblast-conditioned Medium.” The Journal of Cell Biology; 106: 1659-1665. 1988.
[6]  Alliston T, Piek E and Derynck R. The TGF-β family in skeletal development and maintenance. In Derynck R, and Miyazono K. The TGF-β Family. Cold Spring Harbor Press, San Francisco, 2008. 667-723.
[7]  Bonewald LF. “Regulation and regulatory activities of transforming growth factor β.” Crit. Rev. Eukaryot. Gene Expr.; 9: 33-44. 1999.
[8]  Marra F, Bonewald LF, Park-Snyder S, Park IS, Woodruff KA, and Abboud HE. “Characterization and regulation of the latent transforming growth factor-β complex secreted by vascular pericytes.” J. Cell Physiol.; 166: 537-546. 1996.
[9]  Dallas SL, Alliston T, Bonewald LF. Transforming Growth Factor-β. In: Bilezikian JP, Raisz LG, Martin TJ. Principles of Bone Biology, 3rd ed. Academic Press, USA, 2008, 1145-65.
[10]  Taylor AW. “Review of the activation of TGF-β in immunity”. Journal of Leukocyte Biology; 85: 29-35. 2009.
[11]  Annes JP, Munger JS and Rifkin DB. “Making sense of latent TGFb activation”. Journal of Cell Science; 116: 217-224. 2003.
[12]  Assoian RK, Komoriya A, Meyers CA, Miller DM, Sporn MB. “Transforming growth factor-β in human platelets. Identification of a major storage site, purification, and characterization.” J. Biol. Chem.; 258: 7155-7160. 1983.
[13]  Seyedin SM, Thomas TC, Thompson AY, Rosen DM, Piez KA. “Purification and characterization of two cartilage-inducing factors from bovine demineralized bone.” Proc. Natl. Acad. Sci. USA; 82: 2267-227. 1985.
[14]  Janssens K, Dijke P, Janssens S, and Van Hul W. “Transforming growth factor-β 1 to the bone.” Endocr. Rev.; 26, 743-774. 2005.
[15]  Mimura S, Kimura N, Hirata M, Tateyama D, Hayashida M, Umezawa A, Kohara A, Nikawa H, Okamoto T, Furue MK. “Growth factor-defined culture medium for human mesenchymal stem cells.” International Journal of Developmental Biology; 55 (2): 181-187. 2011.
[16]  Jung S, Sen A, Rosenberg L, Behie LA. “Identification of growth and attachment factors for the serum-free isolation and expansion of human mesenchymal stromal cells.” Cytotherapy; 12 (5): 637-657. 2010.
[17]  Vater C, Kasten P, and Stiehler M. “Culture media for the differentiation of mesenchymal stromal cells.” Acta Biomaterialia; 7: 463-477. 2011.
[18]  James D, Levine AJ, Besser D, Hemmati-Brivanlou A. “TGFb activing nodal signaling is necessary for the maintenance of pluripotency in human embryonic stem cells.” Development; 132: 1273-1282. 2005.
[19]  Zhou S. “TGF-β Regulates b-Catenin Signaling and Osteoblast Differentiation in Human Mesenchymal Stem Cells.” Journal of Cellular Biochemistry; 112: 1651-1660. 2011.
[20]  Koli K, Ryynänen MJ, Keski-Oja J. “Latent TGF-β binding proteins (LTBPs)-1 and -3 coordinate proliferation and osteogenic differentiation of human mesenchymal stem cells”. Bone; 43: 679-688. 2008.
[21]  Jian H, Shen X, Liu I, Semenov M, He X, Wang XF. “Smad3-dependent nuclear translocation of β-catenin is required for TGF-β1-induced proliferation of bone marrow-derived adult human mesenchymal stem cells.” Genes Dev; 20: 666-74. 2006.
[22]  DuBose KB, Zayzafoon M and Murphy-Ullrich JE. “Thrombospondin-1 inhibits osteogenic differentiation of human mesenchymal stem cells through latent TGF-b activation.” Biochemical and Biophysical Research Communications; 422: 488-493. 2012.
[23]  Centrella M, Mccarthy TL and Canalis E. “Transforming growth factor β is a bifunctional regulator of replication and collagen synthesis in osteoblast-enriched cell cultures from fetal rat bone”. J. Biol. Chem.; 262: 2869-2874. 1987.
[24]  Pfeilschifte J, D’Souza RSM and Mundy GR. “Effects of transforming growth factor-β on osteoblastic osteosarcoma cells.” Endocrinology; 121: 212-218. 1987.
[25]  Mundy GR and Bonewald LF. “Role of TGF-β in bone remodelling.” Annals New York Academy of Sciences; 593 (1): 97-100. 2006.
[26]  Hock JM, Canalis E, and Centrella M. “Transforming growth factor-β stimulates bone matrix apposition and bone cell replication in cultured fetal rat calvariae.” Endocrinology; 126: 421-426. 1990.
[27]  Chen TL, and Bates RL. “Recombinant human transforming growth factor β-1 modulates bone remodeling in a mineralizing bone organ culture.” J. Bone Miner. Res. 8; 423-434. 1993.
[28]  Takeuchi Y, Nakayama K, and Matsumoto T. “Differentiation and cell surface expression of transforming growth factor-β receptors are regulated by interaction with matrix collagen in murine osteoblastic cells.” J. Biol. Chem.; 271: 3938-3944. 1996.
[29]  Katagiri T, Yamaguchi A, Komaki M, Abe E, Takahashi N, Ikeda T, Rosen V, Wozney JM, Fujisawa-Sehara A, and Suda T. “Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage.” J. Cell Biol.; 127: 1755-1766. 1994.
[30]  Lawrence DA. “Transforming growth factor β: a general review.” Eur. Cytokine Netw; 7: 363-374. 1996.
[31]  Sporn MB, Roberts AB, Wakefield LM, and Crombrugghe BD. “Some Recent Advances in the Chemistry and Biology of Transforming Growth Factor-Β”. The Journal of Cell Biology; 105: 1039-1045. 1987.
[32]  Pertovaara L, Kaipainen A, Mustonen T, Orpana A, Ferrara N, Saksela O and Alitalo K. “Vascular endothelial growth factor is induced in response to transforming growth factor-β in fibroblastic and epithelial cells.” J Biol Chem; 269: 6271-6274. 1994.
[33]  Kay EP, Lee MS, Seong GJ and Lee YG. “TGF-βs stimulate cell proliferation via an autocrine production of FGF-2 in corneal stromal fibroblasts.” Curr Eye Res; 17: 286-293. 1998.
[34]  Brownh PD, Wakefield LM, Levinson AD and Sporn MB. “Physicochemical activation of recombinant latent transforming growth factor-βs 1, 2, and 3.” Growth Factors; 3: 35-43. 1990.
[35]  Khalil N. “TGF-β: from latent to active. Microbes and Infection”; 1: 1255-1263. 1999.
[36]  Barcellos-Hoff MH, Dix TA. “Redox-mediated activation of latent transforming growth factor-β1.” Mol Endocrinol 10: 1079-1083. 1996.
[37]  Schultz-Cherry S, Lawler J, Murphy-Ullrich JE. “The type 1 repeats of thrombospondin 1 activate latent transforming growth factor-b.” J Biol Chem; 269: 26783-26788. 1994.
[38]  Schultz-Cherry S, Chen H, Moser D, Misenheimer TM, Krutzsch HC, Roberts DD, Murphy-Ullrich JE. “Regulation of transforming growth factor-â activation by discrete sequence of thrombospondin-1” J. Biol. Chem.; 270: 7304-7310. 1995.
[39]  Munger JS, Harpel JG, Gleizes P, Mazzieri R, Nunes I, and Rifkin DB. “Latent transforming growth factor-n: Structural features and mechanisms of activation.” Kidney International; 51: 1376-1382. 1997.
[40]  Jullien P, Berg TM, Lawrence DA. “Acidic cellular environments: activation of latent TGF-β and sensitization of cellular responses to TGF-β and EGF.” Int J Cancer.; 43 (5): 886-91. 1989.
[41]  Lawrence DA, Pircher R, and Julien P. “Conversion of a high molecular weight latent [I-TGF from chicken embryo fibroblasts into a low molecular weight active [3-TGF under acidic conditions.” Biochem. Biophys. Res. Commun.; 133: 1026-1034. 1985.
[42]  Brown PD, Wakefield LM, Levinson AD et al. “Physicochemical activation of recombinant latent transforming growth factor-βs 1, 2, and 3.” Growth Factors; 3: 35-43. 1990.
[43]  Moses HL, Shipley GD, Leof EB, Halper J, Coffey RJ, and Tucker RF. “Transforming growth factors. In Control of Animal Cell Proliferation.” In: Boynton AL and Leffert HL, editors. Academic Press, Inc., New York, 1987, 75-92.
[44]  Barcellos-Hoff MH. “Radiation-induced Transforming Growth Factor β and Subsequent Extracellular Matrix Reorganization in Murine Mammary Gland” Cancer Research; 53: 3880-3886. 1993.
[45]  Yang Y, Dignam JD, and Gentry LE. “Role of Carbohydrate Structures in the Binding of β1-Latency-Associated Peptide to Ligands.” Biochemistry; 36: 11923-11932. 1997.
[46]  Vodovotz Y, Chesler L, Chong H, Kim S, Simpson JT, DeGraff W, Cox GW, Roberts AB, Wink DA, and Barcellos-Hoff MH. “Regulation of Transforming Growth Factor β1 by Nitric Oxide.” Cancer Research; 59: 2142-2149. 1999.
[47]  Zamora H, Barclay D, Vodovotz YY. Differential Activation of Recombinant Human Latent Transforming Growth Factor-β1 (Tgf-β1) by acid.” Revista; 15 (2): 177-179. 2007.
[48]  Abe M, Oda N, and Sato Y. “Cell-Associated Activation of Latent Transforming Growth Factor-B By Calpain.” Journal of Cellular Physiology; 174: 186-193. 1998.
[49]  Miyazono K and Heldin C. “Role for carbohydrate structures in TGF-b1 latency.” Nature; 338: 158-60. 1989.
[50]  Munger JS, Huang X, Kawakatsu H, Griffiths MJ, Dalton SL, Wu J, Pittet JF, Kaminski N, Garat C, Matthay MA, Rifkin DB, Sheppard D. “The integrin alphav β 6 binds and activates latent TGF beta 1: a mechanism for regulating pulmonary inflammation and fibrosis.” Cell; 96: 319-328. 1999.
[51]  Breuss JM, Gillett N, Lu L, Sheppard D and Pytela R. “Restricted distribution of integrin β6 mRNA in primate epithelial tissues.” J. Histochem. Cytochem; 41, 1521-1527. 1993.
[52]  Mu D, Cambier S, Fjellbirkeland L, Baron JL, Munger JS, Kawakatsu H, Sheppard D, Broaddus VC and Nishimura SL. “The integrin {alpha}v{β}8 mediates epithelial homeostasis through MT1-MMP-dependent activation of TGF-{β}1.” J. Cell Biol.; 157: 493-507. 2002.
[53]  Knabbe C, Lippman ME, Wakefield LM, Flanders KC, Kasid A, Derynck R, and Dickson RB. “Evidence That Transforming Growth Factor-β is a Hormonally Regulated Negative Growth Factor in Human Breast Cancer Cells.” Cell, Vol. 48, 417-428. 1987.
[54]  Glick AB, Flanders KC, Danielpour D, Yuspa SH, Sporn MB. “Retinoic acid induces transforming growth factor-fl2 in cultured keratinocytes and mouse epidermis.” Cell Regulation; 1: 87-97. 1989.
[55]  Oursler M, Riggs B, Spelsberg T. “Glucocorticoid-induced activation of latent transforming growth factor-b by normal hormone osteoblast-like cell.” Endocrinology; 133: 2187-96. 1993.