Composition of restricted cancer cells which produce cancer cell proliferation suppressive materials, and uses thereof
a cancer cell and suppressive material technology, applied in the direction of tumor/cancer cells, unknown materials, antibody medical ingredients, etc., can solve the problems of host fibrotic response to implant material, limited success, instability of implant material,
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example 1
[0023] This example, and those which follow, employ RENCA cells. These are spontaneous renal adenocarcinoma cells of BALB / C mice, which are widely available, having been maintained in both in vitro cultures and in vivo. See Franco, et al., Cytokine Induced Tumor Immunogenecity, 181-193 (1994).
[0024] Samples of frozen RENCA cells were thawed at 37° C., and then placed in tissue culture flasks containing Dulbecco's Modified Medium (D-MEM), which had been supplemented with 10% bovine serum, penicillin (100 u / ml) and streptomycin (50 ug / ml), to give what will be referred to as “complete medium” hereafter.
[0025] Cells were grown to confluence, and then trypsinized, followed by washing with Hank's Balanced Salt Solution, and then with the complete medium referred to supra.
[0026] In order to determine if the RENCA cells produced tumors efficiently, two BALB / C mice were injected, intraperitoneally, with 106 of these cells. The mice were observed, over a 3-4 week period. Clinically, they ...
example 2
[0028] Following the showing that the RENCA cells did grow in vivo, studies were carried out to determine if these cells grew when restricted in the structure of the invention.
[0029] RENCA cells were grown to confluency, as described supra, trypsinized, and washed, also as described above. Samples of between 60,000 and 90,000 cells were then prepared. The cells were then centrifuged, at 750 RPMs, and fluid was removed. The cells were then suspended in solutions of 1% atelocollagen, in phosphate buffered saline solution, at a pH of 6.5.
[0030] A 1% solution of low viscosity agarose was prepared in minimal essential medium (MEM), maintained at 60° C., and then 100 ul of this was added to the suspension of RENCA cells and atelocollagen, described supra. The materials were then transferred, immediately, as a single large droplet, into sterile, room-temperature mineral oil. The mixture formed a single, smooth, semi-solid bead. This procedure was repeated to produce a number of beads.
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example 3
[0033] Prior to carrying out in vivo investigations, it was necessary to determine if the RENCA cells would grow in beads prepared in the manner described supra.
[0034] To do this, beads prepared as discussed in example 2 were incubated in the medium described in example 2, for a period of three weeks, under the described conditions. Three of the beads were then cut into small pieces, and cultured in standard culture flasks, affording direct contact with both the flask and culture medium.
[0035] Observation of these cultures indicated that the cells grew and formed standard RENCA colonies. This indicated that the cells had remained viable in the beads.
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