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Density-dependent growth regulation of pig retinal pigment epithelial cells in vitro

  • Tongalp H. Tezel
  • Lucian V. Del Priore
Vitreo-Retinal

Abstract

• Background: Transplantation of the retinal pigment epithelium (RPE) involves delivering donor RPE onto a bare area of host Bruch's membrane that is surrounded by a confluent monolayer of RPE. We investigated the effects of different plating densities and the presence of an adjacent confluent monolayer on the growth characteristics and final morphology of pig RPE in vitro. • Methods: In single-well experiments, porcine RPE were plated in 24-well plates at densities varying from 1 to 75 cells/mm2. Triplicate plates were counted on the 3rd and 10th days after plating and at confluence. A multiwell chamber was built to allow cells plated at different densities to be bathed with conditioned media from adjoining wells. • Results: In single-well experiments, plating at low densities increased the time to reach confluence and resulted in fewer, larger and more fusiform RPE at confluence. In multiwell experiments, the growth rate of cells plated at low density decreased as the amount of high-density medium increased in communicating wells and led to smaller, rounder cells at confluence. The presence of low-density RPE in adjoining wells increased the growth rate of RPE plated at high density and produced fewer, larger and more fusiform cells at confluence. Newly plated RPE grew more slowly when confluent monolayers of RPE were present in adjoining wells. • Conclusions: Plating density is a critical factor in determining the growth rate and the final morphology of RPE in tissue culture. The presence of a neighboring confluent monolayer of RPE inhibits the growth rate of newly plated RPE in vitro.

Keywords

Growth Rate Growth Regulation Conditioned Medium Retinal Pigment Epithelium Critical Factor 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Adelberg DA, Del Priore LV, Kaplan HJ (1995) Surgical excision of idiopathic and miscellaneous subfoveal membranes (SFNVM) from myopia, angioid streaks and miscellaneous disorders. Retina 15:298–305Google Scholar
  2. 2.
    Berger AS, Kaplan HJ (1992) Clinical experience with surgical removal subfoveal neovascular membranes. Ophthalmology 99:969–976PubMedGoogle Scholar
  3. 3.
    Bost LM, Aootaki-keen AE, Hjelmeland LM (1992) Coexpression of FGF-5 and bFGF by the retinal pigment epithelium in vitro. Exp Eye Res 55:727–734CrossRefPubMedGoogle Scholar
  4. 4.
    Burke BM (1989) Stimulation of DNA synthesis in human and bovine RPE by peptide growth factors, the response to TNFα and EGF is dependent upon culture density. Curr Eye Res 12:1278–1286Google Scholar
  5. 5.
    Campochiaro PA, Hackett SF, Conway BP (1991) Retinoic acid promotes density-dependent growth arrest in human retinal pigment epithelial cells. Invest Ophthalmol Vis Sci 32:65–72PubMedGoogle Scholar
  6. 6.
    Cremers FPM, van de Pol DIRK, van der Kerkhoff PM, Wieringa B, Ropers HH (1990) Cloning a gene that is rearranged in patients with cboroideremia. Nature 347:674–677CrossRefPubMedGoogle Scholar
  7. 7.
    Dawson-Saunders B, Trapp RG (1994) Basic and clinical biostatistics, 2nd edn. Appleton and Lange, Norwalk, ConnGoogle Scholar
  8. 8.
    Del Priore LV, Kaplan HJ, Silvermann MS, Valentino T, Mason G, Hornbeck R (1993) Experimental and surgical aspects of retinal pigment epithelial cell transplantation. Eur J Implant Ref Surg 5:128–132Google Scholar
  9. 9.
    Del Priore LV, Hornbeck R, Kaplan HJ, Jones Z, Valentino T, Silverman MS, Mosinger-Ogilvie J, Swinn M (1995) Debridement of the pig retinal pigment epithelium in vivo. Arch Ophthalmol 113:939–944PubMedGoogle Scholar
  10. 10.
    Desai VN, Del Priore LV, Kaplan HJ (1994) Choriocapillaris atrophy after submacular surgery in the presumed ocular histoplasmosis syndrome. Arch Ophthalmol 113:409–410Google Scholar
  11. 11.
    Froesch ER, Schimid C, Schwander J, Zapf J (1985) Actions of insulin-like growth factors. Ann Rev Physiol 47:443–467Google Scholar
  12. 12.
    Grierson I, Hiscott P, Hogg P, Robey H, Mazure A, Larkin G (1994) Development, repair and regeneration of the retinal pigment epithelium. Eye 8:255–262PubMedGoogle Scholar
  13. 13.
    Holley RW (1975) Control of growth of mammalian cells in cell culture. Nature 258:487–490CrossRefPubMedGoogle Scholar
  14. 14.
    Kishi H, Mishima HK, Yamashita U (1994) Growth regulation of retinal pigment epithelial cell in vitro. Curr Eye Res 13:661–668PubMedGoogle Scholar
  15. 15.
    Kitaoka T, Bost LM, Ishigooka H, Aootaki-keen AE, Hjelmeland LM (1993) Increasing cell density downregulates the expression of acidic FGF by human RPE cells in vitro. Curr Eye Res 12:993–999PubMedGoogle Scholar
  16. 16.
    Korte GE, Reppuci V, Henkind P (1984) RPE destruction causes choriocapillaris atrophy. Invest Ophthalmol Vis Sci 25: 1135–1145PubMedGoogle Scholar
  17. 17.
    Mitchell GA, Brody LC, Sipila I, Looney JE, Wong C, Engelhardt JF, Patel AS, Steel G, Obie C, Kaiser-Kupfer M (1989) At least two mutant alleles of ornithine-d-aminotrans-ferase cause gyrate atrophy of the choroid and retina in Finns. Proc Natl Acad Sci USA 86:197–201PubMedGoogle Scholar
  18. 18.
    Nasir M, Zarbin MA (1993) Choriocapillaris atrophy as a complication of surgical excision of choroidal neovascular membranes. Invest Ophthalmol Vis Sci 34 [Suppl]:834Google Scholar
  19. 19.
    Phillips HJ (1973) Dye exclusion tests for cell viability. In: Krause PF, Patterson MK Jr (eds) Tissue culture: methods and applications. Academic Press, New York, pp 406–408Google Scholar
  20. 20.
    Pollack JS, Kaplan HJ, Del Prime LV, Smith ME (1993) Choriocapillaris atrophy associated with age-related macular degeneration. Invest Ophthalmol Vis Sci 34 [Suppl]:834Google Scholar
  21. 21.
    Pollack JS, Kaplan HJ, Del Priore LV, Smith ME (1993) Choriocapillaris atrophy following subfoveal membrane excision in exudative age-related macular degeneration. Ophthalmology 100 [Suppl]:122Google Scholar
  22. 22.
    Song M, Lui GM (1990) Propagation of fetal human RPE cells: preservation of original culture morphology after serial passage. J Cell Physiol 143:196–203CrossRefPubMedGoogle Scholar
  23. 23.
    Zarbin MA, Nasir M (1993) Impaired choriocapillaris perfusion following subfoveal surgery for macular degeneration. Ophthalmology 100 [Suppl]:97Google Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Tongalp H. Tezel
    • 1
  • Lucian V. Del Priore
    • 1
  1. 1.Department of Ophthalmology & Visual SciencesWashington University School of MedicineSaint LouisUSA

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