Hostname: page-component-7479d7b7d-68ccn Total loading time: 0 Render date: 2024-07-12T06:25:35.671Z Has data issue: false hasContentIssue false
  • English
  • Français

Generation of Differentiating Human Epidermal Cells on Basement Membrane Matrigel: Potential for use as Autografts in Recurrent Ulcerative Lesions

Published online by Cambridge University Press:  26 February 2011

Dorothy G. Walker-Jones ,
C. A. Reindorf ,
E. Massac Jr ,
A. D. Adekile ,
M. G. Hall Jr  and
A. I. Nunez
Dorothy G. Walker-Jones
Affiliation:
Howard University, Departments of Zoology and the The Sickle Cell Center, 2400 Sisxth St., N. W., Washington, D. C. 20059
C. A. Reindorf
Affiliation:
and The Sickle Cell Center, 2400 Sixth St., N. W., Washington, D. C. 20059
E. Massac Jr
Affiliation:
and Surgery and The Sickle Cell Center, 2400 Sixth St., N. W., Washington, D. C. 20059
A. D. Adekile
Affiliation:
University of Ife, Department of Pediatrics and Chilc Health, Ife-Ife, Oyo State, Nigeria
M. G. Hall Jr
Affiliation:
and Surgery and The Sickle Cell Center, 2400 Sixth St., N. W., Washington, D. C. 20059
A. I. Nunez
Affiliation:
Howard University, Departments of Zoology and the The Sickle Cell Center, 2400 Sisxth St., N. W., Washington, D. C. 20059
Get access

Abstract

The successful use of in vitro-generated skin autografts in the treatment of burns in humans (Green and O'Connor, 1984) has advanced the potential for application in the treatment of recurrent ulcerative lesions that occur in some patients with sickle cell disease and other disorders. Conventional grafts frequently break down in these patients, requiring repeated grafting that may eventually result in the lack of suitable donor sites. Preliminary studies show that Type I collagen (Collistat) promotes healing of sickle cell-lesions but with extensive scarring and loss of pigmentation (Adekile et al., 1987). Therefore, the primary aim of this study was the development of a culture procedure for the rapid generation of differentiating human epidermal cells on a biomatrix for use in conjunction with the Collistat, and later, the establishment of a frozen culture bank. Dispasefollowed by trypsin-dissociated inner epidermal cells from dermatome sections of skin of adult patients undergoing elective plastic surgery were cultured on basement membrane-derived Matrigel (Collaborative Research, Inc.) coated onto plastic coverslips. Growth was established in RPMI1640 or IMEM supplemented with insulin, transferrin, hydrocortisone and bovine pituitary extract or Keratinocyte Growth Medium (KGM)a nd from 0.5–2.0% fetal bovine serum. The epidermal cells exhibited active migration and the formation of bi- and tri-layered aggregates by day 3. Large multilayered aggregates were produced in KGM with 1.16 mM CaCl2 added. Electron microscopic observations revealed increased numbers of desmosomal junctions, interdigitations and apical-basal polarity in these aggregates. A distinguishing feature ofthese and other cells grown on Matrigel is the absence of the degree of spreading that is characteristic of cells grown on plastic. It is noteworthy that the epidermal cells did not invade through the Matrigel.

Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 110: Symposium M – Biomedical Materials and Devices , 1987 , 393
Copyright
Copyright © Materials Research Society 1988

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1. Adekile, A. D., Reindorf, C. A., Walker-Jones, D., Report to the US Agency for International Development, 1987 (unpublished).Google Scholar
2. Aresu, O., Schmidt, J., Agostacchio, C., Allevena, G., Melchiori, A., Parodi, S., Albini, A., J. Cell Biol. 105, 220a, #1242 (1987).Google Scholar
3. Boyce, S. T. and Ham, R. G., J. Invest. Dermat. 81, 33s (1983); In Vitro Models for Cancer Res, III, 245 (1986).Google Scholar
4. Eisinger, M., Monden, M., Raaf, J. H., Fortner, J. G., Surgery 80, 287 (1980).Google Scholar
5. Green, H., Kehinde, O., Thomas, J., Proc. Nat. Acad. Sci. USA 76, (5), 665 (1979).Google Scholar
6. Green, H. and O'Connor, N., in Burn Wound Coverin, edited by Wise, D. (CRC Press, Inc., Boca Raton, Fla., 1984), p. 39.Google Scholar
7. Kleinman, H. K., McGarvey, M. L., Liotta, L. A., Robey, P. G., Tryggvason, K., Martin, G. R., Biochemistry 21, 6188 (1982).CrossRefGoogle Scholar
8. Moscona, A., Expl. Cell Res. 22, 455 (1961).Google Scholar
9. O'Connor, N., Mulliken, J. B., Banks-Schlegel, S., Kehinde, O., Green, H., The Lancet 1, 75 (1981).CrossRefGoogle Scholar
10. Valentich, J. D. and Garretson, L. T., J. Cell Biol. 105, 140a, #787 (1987).Google Scholar
11. Watt, F. M. and Green, H., Nature 295, 434 (1982).Google Scholar
12. Yannas, I. V. and Burke, J. F., Harvard Mag. Sept.-Oct., 1 (1984).Google Scholar