Splitting culture medium by air-jet and rewetting for the assessment of the wettability of cultured epithelial cell surfaces.

This study found that the phenomenon of rewetting after squeezing culture medium varied in different culture conditions for rat oral mucosal epithelial cells. When culture medium covering over cultured cells was squeezed by an air-jet application, the motion of squeezed culture medium was able to be observed by using a commercially available movie camera. Squeezed width on cells cultured in keratinocyte culture medium (KCM), which contained with fetal bovine serum, was one-sixth of that in FBS-free KCM. This result corresponded to the mucous layer staining statuses of cultured cells in both cases; positive in KCM and negative in FBS-free medium. Furthermore, the gene expression of mucous glycoprotein MUC4 in KCM was 100 times higher than that in FBS-free medium, and the expression of MUC4 protein only showed on the apical surface of cells cultured in KCM. The relative gene expression levels of MUC1, 13, 15, and 16 in both the normal and FBS-free medium were found to be no more than one-thirtieth of that of MUC4 in KCM. The main factor of the wettability difference between KCM and FBS-free medium was speculated to be the difference of MUC4 expression between both media. This method can be a simple technique for testing not only the surface wettability but also the mucous formation of cultured cells.

[1]  C. Dawson,et al.  Demonstration of the mucous layer of the tear film by electron microscopy. , 1985, Investigative ophthalmology & visual science.

[2]  A. Allen,et al.  Gastroduodenal mucus bicarbonate barrier: protection against acid and pepsin. , 2005, American journal of physiology. Cell physiology.

[3]  M. Lemp,et al.  Wettability and wetting of corneal epithelium. , 1971, Experimental eye research.

[4]  T. Okano,et al.  Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium. , 2004, The New England journal of medicine.

[5]  T. Okano,et al.  Thermo‐responsive polymeric surfaces; control of attachment and detachment of cultured cells , 1990 .

[6]  Christophe Baudouin,et al.  The definition and classification of dry eye disease: report of the Definition and Classification Subcommittee of the International Dry Eye WorkShop (2007). , 2007, The ocular surface.

[7]  Rama Bansil,et al.  Mucin structure, aggregation, physiological functions and biomedical applications , 2006 .

[8]  H. Green,et al.  Seria cultivation of strains of human epidemal keratinocytes: the formation keratinizin colonies from single cell is , 1975, Cell.

[9]  M. Yamato,et al.  Differential expression of MUC16 in human oral mucosal epithelium and cultivated epithelial sheets. , 2007, Experimental eye research.

[10]  N. Koizumi,et al.  Establishment of a cultivated human conjunctival epithelium as an alternative tissue source for autologous corneal epithelial transplantation. , 2006, Investigative ophthalmology & visual science.

[11]  Hideo Namiki,et al.  The effect of micropores in the surface of temperature-responsive culture inserts on the fabrication of transplantable canine oral mucosal epithelial cell sheets. , 2006, Biomaterials.

[12]  Teruo Okano,et al.  Tissue engineered myoblast sheets improved cardiac function sufficiently to discontinue LVAS in a patient with DCM: report of a case , 2012, Surgery Today.

[13]  Hideo Namiki,et al.  Prevention of esophageal stricture after endoscopic submucosal dissection using tissue-engineered cell sheets ( , 2013 .

[14]  K. Carraway,et al.  Mucin Structure and Function: Insights from Molecular Biology , 1995 .

[15]  I. Gipson,et al.  Differential regulation of membrane-associated mucins in the human ocular surface epithelium. , 2004, Investigative ophthalmology & visual science.

[16]  E. Hounsell,et al.  O-linked protein glycosylation structure and function , 1996, Glycoconjugate Journal.

[17]  T. Okano,et al.  The regulation of epithelial cell proliferation and growth by IL-1 receptor antagonist. , 2013, Biomaterials.

[18]  M. Nakamura,et al.  Improvement of corneal barrier function by the P2Y(2) agonist INS365 in a rat dry eye model. , 2001, Investigative ophthalmology & visual science.

[19]  Masayuki Yamato,et al.  Functional bioengineered corneal epithelial sheet grafts from corneal stem cells expanded ex vivo on a temperature-responsive cell culture surface , 2004, Transplantation.

[20]  L. Holm,et al.  The adherent gastrointestinal mucus gel layer: thickness and physical state in vivo. , 2001, American journal of physiology. Gastrointestinal and liver physiology.

[21]  Henry D Perry,et al.  Dry eye disease: pathophysiology, classification, and diagnosis. , 2008, The American journal of managed care.

[22]  H Green,et al.  Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. , 1975, Cell.

[23]  T. Okano,et al.  Cell sheet engineering for myocardial tissue reconstruction. , 2003, Biomaterials.

[24]  T. Okano,et al.  Thermo-responsive culture dishes allow the intact harvest of multilayered keratinocyte sheets without dispase by reducing temperature. , 2001, Tissue engineering.

[25]  S. Feinberg,et al.  Development and Characterization of a Tissue-engineered Human Oral Mucosa Equivalent Produced in a Serum-free Culture System , 2000, Journal of dental research.

[26]  J. Daniels,et al.  Autologous serum eyedrops for dry eyes and epithelial defects: clinical and in vitro toxicity studies , 2001, The British journal of ophthalmology.

[27]  A. Vigorito,et al.  GVHD dry eyes treated with autologous serum tears , 2000, Bone Marrow Transplantation.

[28]  T. Okano,et al.  Fabrication of transplantable human oral mucosal epithelial cell sheets using temperature-responsive culture inserts without feeder layer cells , 2006, Journal of Artificial Organs.

[29]  Imin Kao,et al.  Noncontact Active Sensing for Viscoelastic Parameters of Tissue With Coupling Effect , 2011, IEEE Transactions on Biomedical Engineering.

[30]  K. Tsubota,et al.  Treatment of dry eye by autologous serum application in Sjögren’s syndrome , 1998, The British journal of ophthalmology.

[31]  T. Okano,et al.  Preparation of keratinocyte culture medium for the clinical applications of regenerative medicine , 2011, Journal of Tissue Engineering and Regenerative Medicine.

[32]  A. Hoffman,et al.  Quantitation of rabbit corneal epithelial cell outgrowth on polymeric substrates in vitro. , 1990, Investigative ophthalmology & visual science.

[33]  Masayuki Yamato,et al.  Periodontal regeneration with multi-layered periodontal ligament-derived cell sheets in a canine model. , 2009, Biomaterials.

[34]  Yukihiro Matsumoto,et al.  A new noninvasive tear stability analysis system for the assessment of dry eyes. , 2004, Investigative ophthalmology & visual science.

[35]  S. Kinoshita,et al.  Barrier Function of Ocular Surface Epithelium , 1997 .

[36]  H. Shinohara,et al.  Rebamipide increases the mucin-like glycoprotein production in corneal epithelial cells. , 2012, Journal of ocular pharmacology and therapeutics : the official journal of the Association for Ocular Pharmacology and Therapeutics.

[37]  B. Deplancke,et al.  Microbial modulation of innate defense: goblet cells and the intestinal mucus layer. , 2001, The American journal of clinical nutrition.

[38]  Mitchell P. Weikert,et al.  Noninvasive assessment of tear stability with the tear stability analysis system in tear dysfunction patients. , 2011, Investigative ophthalmology & visual science.

[39]  R. N. Wenzel.  RESISTANCE OF SOLID SURFACES TO WETTING BY WATER , 1936 .