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Human dermal reticular fibroblasts at confluence display a signature micro pattern in vitro

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DOI: 10.4236/ojrm.2013.24014    5,087 Downloads   8,249 Views   Citations

ABSTRACT

This paper sets out to demonstrate that scraping of the flat dorsal surface of human dermis with a scalpel blade and cell plating without centrifugation can lead to the recognition and identification of the individual packing micro pattern of dermal reticular fibroblasts at confluence. The characteristic alignment of papillary and reticular fibroblasts at right angles to each other led to the positive identification of reticular fibroblasts. A non-enzymatic means of sub-culturing (passaging), which yields fully functional, healthy cells with normal, phenotypic morphology is also described. Implications for published subcutaneous wound healing studies are discussed as well as the confluent reticular fibroblast configuration, interpreted as ananatomic site identity code,which may be the address of a specific fibroblast gene pattern expression.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Solomon, D. (2013) Human dermal reticular fibroblasts at confluence display a signature micro pattern in vitro. Open Journal of Regenerative Medicine, 2, 99-105. doi: 10.4236/ojrm.2013.24014.

References

[1] MacNeil, S. (2007) Progress and opportunities for tissueengineered skin. Nature, 445, 874-880.
http://dx.doi.org/10.1038/nature05664
[2] McDougall, S., Dallon, J., Sherratt, J. and Maini, P. (2006) Fibroblast migration and collagen deposition during dermal wound healing: Mathematical modelling and clinical implications. Philosophical Transactions of the Royal Society A, 364, 1385-1405.
http://dx.doi.org/10.1098/rsta.2006.1773
[3] Sorrell, J.M. and Caplan, A.I. (2004) Fibroblast heterogeneity: More than skin deep. Journal of Cell Science, 117, 667-675. http://dx.doi.org/10.1242/jcs.01005
[4] Sorrell, J.M., Baber, M.A. and Caplan, A.I. (2004) Sitematched papillary and reticular human dermal fibroblasts differ in their release of specific growth factors/cytokines and in their interaction with keratinocytes. Journal of Cellular Physiology, 200, 134-145.
http://dx.doi.org/10.1002/jcp.10474
[5] Solomon, D.E. (2012) Mimicry of a natural, living intraepidermal micro pattern used in guided tissue regeneration of the human epidermis. Journal of Developmental Biology and Tissue Engineering, 4, 1-7.
[6] Normand, J. and Karasek, M.A. (1995) A method for the isolation and serial propagation of keratinocytes, endothelial cells, and fibroblasts from a single punch biopsy of human skin. In Vitro Cellular & Developmental Biology: Animals, 31, 447-455.
http://dx.doi.org/10.1007/BF02634257
[7] Pandamooz, S., Hadipour, A., Akhavan-Niaki, H., Pourghasem, M., Abedian, Z., Ardekani, A.M., Golpour, M., Hassan, Z.M. and Mostafazadeh A. (2012) Short exposure to collagenase and coculture with mouse embryonic pancreas improve human dermal fibroblast culture. Biotechnology and Applied Biochemistry, 59, 254-261.
http://dx.doi.org/10.1002/bab.1020
[8] Pageon, H., Zucchi, H. and Asselineau, D. (2012) Distinct and complementary roles of papillary and reticular fibroblasts in skin morphogenesis and homeostasis. European Journal of Dermatology, 22, 324-332.
[9] Sandulache, V.C., Zhou, Z., Sherman, A., Dohar, J.E. and Hebda, P.A. (2003) Impact of transplanted fibroblast on rabbit skin wounds. JAMA Otolaryngology: Head & Neck Surgery, 129, 345-350.
http://dx.doi.org/10.1001/archotol.129.3.345
[10] Sch?nherr, E., Beavan, L.A., Hausser, H., Kresse, H. and Culp, L.A. (1993) Differences in decorin expression by papillary and reticular fibroblasts in vivo and in vitro. Biochemical Journal, 290, 893-899.
[11] Munavalli, G.S., Smith, S., Maslowski, J.M. and Weiss, R.A. (2013) Successful treatment of depressed, distensible acne scars using autologous fibroblasts: A multi-site, prospective, double blind, placebo-controlled clinical trial. Dermatologic Surgery, 39, 1226-1236.
http://dx.doi.org/10.1111/dsu.12204
[12] Solomon, D.E. (2002) An in vitro examination of an extracellular matrix scaffold for use in wound healing. International Journal of Experimental Pathology, 83, 209-216. http://dx.doi.org/10.1046/j.1365-2613.2002.00238.x
[13] Korner, G., Bjornsson, T.D. and Vlodavsky, I. (1993) Extracellular matrix produced by cultured corneal and aortic endothelial cell contains active tissue type and urokinasetype plasminogen activators. Journal of Cellular Physiology, 154, 456-465.
http://dx.doi.org/10.1002/jcp.1041540303
[14] Rosen, J.J. and Culp, L.A. (1977) Morphology and cellular origins of substrate-attached material from mouse fibroblasts. Experimental Cell Research, 107, 139-150.
http://dx.doi.org/10.1016/0014-4827(77)90395-0
[15] Kramer, R.H., Gonzalez, R. and Nicholson, G.L. (1980) Metastatic tumor cells adhere preferentially to the extracellular matrix underlying vascular endothelial cells. International Journal of Cancer, 26, 639-645.
http://dx.doi.org/10.1002/ijc.2910260516
[16] Li, J., Thielemann, C., Reuning, U. and Johannsmann, D. (2005) Monitoring of integrin-mediated adhesion of human ovarian cancer cells to model protein surfaces by quartz crystal resonators: Evaluation in the impedance analysis mode. Biosensors and Bioelectronics, 20, 1333-1340. http://dx.doi.org/10.1016/j.bios.2004.05.004
[17] Voyta, J.C., Via, D.P., Butterfield, C.E. and Zetter, B.R. (1984) Identification and isolation of endothelial cells based on their increased uptake of acetylated low density lipoprotein. Journal of Cell Biology, 99, 2034-2040.
http://dx.doi.org/10.1083/jcb.99.6.2034
[18] Sutherland, J., Denyer, M. and Britland, S. (2005) Con-tact guidance in human dermal fibroblasts is modulated by population pressure. Journal of Anatomy, 206, 581-587. http://dx.doi.org/10.1111/j.1469-7580.2005.00415.x
[19] Sorrell, J.M. and Caplan, A.I. (2009) Fibroblasts—A diverse population at the center of it all. International Review of Cell and Molecular Biology, 276, 161-214.
http://dx.doi.org/10.1016/S1937-6448(09)76004-6
[20] Brem, H., Golinko, M.S., Stojadinovic, O., Kodra, A., Diegelmann, R.F., Vukelic, S., Entero, H., Coppock, D.L. and Tomic-Canic, M. (2008) Primary cultured fibroblasts derived from patients with chronic wounds: A methodology to produce human cell lines and test putative growth factor therapy such as GMCSF. Journal of Translational Medicine, 6, 75.
http://dx.doi.org/10.1186/1479-5876-6-75
[21] Brem, H., Stojadinovic, O., Diegelmann, R.F., Entero, H., Lee, B., Pastar, I., Golinko, M., Rosenberg, H. and Tomic-Canic, M. (2007) Molecular markers in patients with chronic wounds to guide surgical debridement. Molecular Medicine, 1, 30-39.
http://dx.doi.org/10.2119/2006-00054.Vittorini
[22] Egles, C., Shamis, Y., Mauney, J.R., Volloch, V., Kaplan, D.L. and Garlick, J.A. (2008) Denatured collagen modulates the phenotype of normal and wounded human skin equivalents. Journal of Investigative Dermatology, 128, 1830-1837. http://dx.doi.org/10.1038/sj.jid.5701240
[23] Vogel, K.G. (1978) Effects of hyaluronidase, trypsin, and EDTA on surface composition and topography during detachment of cells in culture. Experimental Cell Research, 113, 345-357.
http://dx.doi.org/10.1016/0014-4827(78)90375-0
[24] Sorrell, J.M., Baber, M.A., Caplan, A.I. (2006) Clonal characterization of fibroblasts in the superficial layer of the adult human dermis. Cell and Tissue Research, 327, 499-510. http://dx.doi.org/10.1007/s00441-006-0317-y
[25] Clark, R.A.F. (1996) Wound repair: Overview and general considerations. In: Clark, R.A.F., Ed., The Molecular, Cellular Biology of Wound Repair, Plenum Press, New York, 3-35.
[26] Singer, A.J. and Clark, R.A.F. (1999) Cutaneous wound healing. The New England Journal of Medicine, 341, 738-746. http://dx.doi.org/10.1056/NEJM199909023411006
[27] Sorrell, J.M. and Caplan, A.I. (2010) Topical delivery of mesenchymal stem cells and their function in wounds. Stem Cell Research & Therapy, 1, 30.
http://dx.doi.org/10.1186/scrt30
[28] Rinn, J.L., Bondre, C., Gladstone, H.B., Brown, O., Chang, H.Y. (2006) Anatomic demarcation by positional variation in fibroblast gene expression. PLoS Genetics, 2, e119.
http://dx.doi.org/10.1371/journal.pgen.0020119
[29] Janson, D.G., Saintigny, G., Van Adrichem, A., Mahé, C. and El-Ghalbzouri, A. (2012) Different gene expression patterns in human papillary and reticular fibroblasts. Journal of Investigative Dermatology, 132, 2565-2572.
http://dx.doi.org/10.1038/jid.2012.192
[30] Watson, D., Keller, G.S., Lacombe, V., Fodor, P.B., Rawnsley, J., Lask, G.P. (1999) Autologous fibroblasts for treatment of facial rhytids and dermal depressions. A pilot study. Archives of Facial Plastic Surgery, 1, 165-170.
http://dx.doi.org/10.1001/archfaci.1.3.165
[31] Li, W., Fan, J., Chen, M., Guan, S., Sawcer, D., Bokoch, G.M. and Woodley, D.T. (2004) Mechanism of human dermal fibroblast migration driven by Type 1 collagen and platelet-derived growth factor-BB. Molecular Biology of the Cell, 15, 294-309.
http://dx.doi.org/10.1091/mbc.E03-05-0352
[32] Novotny, G.E.K. and Gnoth, C. (1991) Variability of fibroblast morphology in vivo: A silver impregnation study on human digital dermis and subcutis. Journal of Anatomy, 177, 195-207.
[33] Mine, S., Fortunel, N.O., Pageon, H. and Asselineau, D. (2008) Aging alters functionally human dermal papillary fibroblasts but not reticular fibroblasts: A new view of skin morphogenesis and aging. PLoS One, 3, e4066.
http://dx.doi.org/10.1371/journal.pone.0004066
[34] Solomon, D.E. (2011) An in vitro human skin assay protocol which directly uses the natural cellular interactions between unmanipulated autologous human epidermal and dermal cells. United Kingdom Patent Gazette (Intellectual Property Office), GB2442271B.

  
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