Lesional psoriatic kerationocyte (LPK) culture is considered to be difficult under high-Ca2+ conditions in the absence of special proliferative agents. Using a permeable collagen membrane, we obtained a culture of LPKs under high-Ca2+ conditions without any special proliferative agents. Single-cell suspensions were prepared from the epidermis of chronic psoriatic plaques. Cells were inoculated on the collagen membrane suspended slightly above the bottom of a Petri dish. We used a culture medium of Eagle's MEM containing 10% fetal calf serum. LPKs attached to the membrane 12 h after inoculation and gradually spread. They reached a confluent state by the 10th day of culture. We measured the concentrations of TGFα and IL-6 in the medium of LPKs, and compared these with the concentrations in normal keratinocyte (NK) cultures. Significantly increased secretion of TGFα by LPKs was observed during the initial phase but this secretion subsequently decreased. Concentrations of IL-6 were below the detectable level in both of NKs and LPKs throughout the observation period. Our results demonstrate that cultured LPKs under high-Ca2+ conditions secrete a larger amount of TGFα but not IL-6. Our cell culture system, which allows LPKs to spread and stratify, contributes to the study of the pathogenesis of psoriasis.
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Nakayama J, Yasumoto S, Takagi Y, Nagae S, Yashima Y, Urabe H (1983) Studies of cell and organ culture of psoriatic and normal keratinocytes in vitro. J Dermatol 10: 411–419
Nickoloff BJ, Mitra RS, Elder JT, Fisher GJ, Voorhees JJ (1989) Decreased growth inhibition by recombinant gamma interferon is associated with increased transforming growth factor-α production in keratinocytes cultured from psoriatic lesions. Br J Dermatol 121: 161–174
Baden HP, Kubilus J, MacDonald MJ (1981) Normal and psoriatic keratinocytes and fibroblasts compared in culture. J Invest Dermatol 76: 53–55
Hennings H, Michael D, Cheng C, Steinert P, Holbrook K, Yuspa SH (1980) Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell 19: 245–254
Mitsuhashi Y, Mikami Y, Mikami H, Ishikawa H, Tamai K, Hashimoto I (1993) Simultaneous and separated culture of keratinocytes and fibroblasts on each side of collagen membrane. J Dermatol Sci 5: 3–13
Yoshizato K, Nishikawa A, Taira T (1988) Functionally polarized layers formed by epidermal cells on a permeable transport collagen film. J Cell Sci 91: 491–499
Lisby S, Ralfkiaer E, Rothlein R, Vejlsgaard GL (1989) Intercellular adhesion molecule-1 (ICAM-1) expression correlated to inflammation. Br J Dermatol 120: 479–484
Coffey RJ, Derynck R, Wilcox JN, Bringman TS, Goustin AS, Moses HL, Pittelkow MR (1987) Production and auto-induction of transforming growth factor-α in human keratinocytes. Nature 328: 817–820
Gottlieb AB, Chang CK, Posnett DN, Fanelli B, Tam JP (1988) Detection of transforming growth factor α in normal, malignant, and hyperproliferative human keratinocytes. J Exp Med 167: 670–675
Elder JT, Fisher GJ, Lindquest PB, Bennertt GL, Pittelkow MR, Coeffey RJ, Ellingsworth L Jr, Derynck R, Voorhees JJ (1989) Over expression of transforming growth factor α in psoriatic epidermis. Science 243: 811–814
Nanney LB, Yates RA, King LE Jr (1992) Modulation of epidermal growth factor receptors in psoriatic lesions during treatment with topical EGF. J Invest Dermatol 98: 296–301
Grossman RM, Krueger J, Yourish D, Granelli-Piperno A, Murphy DP, May LT, Kupper TS, Sehgel PB, Gottlieb AB (1989) Interleukin 6 is expressed in high levels in psoriatic skin and stimulates proliferation of cultured human keratinocytes. Proc Natl Acad Sci USA 86: 6367–6371
Prens EP, Benne K, Damma JV, Bakkus M, Brakel K, Benner R, Joost TV (1990) Interleukin-1 and interleukin-6 in psoriasis. J Invest Dermatol 95: 121s-124s
Gillitzer R, Berger R, Mielke V, Mueller C, Wolff K, Stingl G (1991) Upper keratinocytes of psoriatic lesions express high levels of NAP-1/IL-8. J Invest Dermatol 97: 73–79
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Miyauchi, Y., Mitsuhashi, Y., Kikuchi, T. et al. Feasibility of in vitro culturing of lesional psoriatic keratinocytes in medium containing high calcium concentrations. Arch Dermatol Res 287, 731–734 (1995). https://doi.org/10.1007/BF01105797
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