• Archives of Plastic Surgery
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• v.35 no.4
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• pp.385-392
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• 2008

Purpose: The most effective methods of harvesting, preparing, and injecting autologous fat grafts have been inconsistent and conflicting. With its limitation as resorption in fat grafting, handling various techniques affect adipocyte survival, and is crucial to optimizing its long-term survival. To improve graft survival, re-implantation of cryopreserved adipocytes was developed. In addition, adipocytes do not induce immune rejection in response to non-self lymphocytes in a mixed lymphocyte reaction. The purpose of this study is to analyze the changes in cryopreserved adipocytes so as to determine the most efficient long-term storage period, and to analyze the changes in cryopreserved allografted adipocytes so as to determine the efficacy of cryopreserved adipocytes allografting. Methods: Fat tissues were harvested from the inguinal and retroperitoneal fat pad of mice. After the centrifugation of the harvested fat tissues, they were disintegrated with collagenase. The adipocytes were obtained by centrifugation of the disintegrated fat tissues. The adipocytes were treated as follows: (1) They were examined for weight and then frozen at $-20^{\circ}C$(n=25). For four months, each five frozen samples were taken and examined for weight and histologic changes in the 1st week, the 1st month, the 2nd month, the 3rd month, and the 4th month, respectively. (2) The adipocytes were immediately frozen at $-20^{\circ}C$(n=125). For four months, five frozen samples were taken, and allografted in the same time period as above. Finally, for four months, five cryopreserved allografted adipocytes were taken and examined for histologic changes in the same time period as above. Results: (1) Significant weight changes and histologic findings with inflammatory and destructive changes were observed in the cryopreserved adipocytes in three months. (2) Significant fat necrotic changes in the histologic changes with Hematoxylin and eosin stain were observed in the cryopreserved allografted adipocytes since the first week, independent of the freezing period. Conclusion: The study results show that the adipocytes that were cryopreserved for more than three months underwent obvious weight reductions and necrotic changes, and the adipocytes that were allografted without freezing were viable for four months, but the cryopreserved allografted adipocytes had obvious necrotic changes since the first week regardless of the freezing period.

• Korean Journal of Pharmacognosy
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• v.24 no.4
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• pp.267-281
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• 1993

To find antitumor components from higher fungi, the mycelia of Collybia confluens (Pers. ex Fr.) Kummer were cultured in artificial media. For efficient production of the mycelia, the influences of various modifications of culture conditions were examined. A water-soluble protein-bound polysaccharide fraction, Fr. A, was obtained from the mycelia by hot water extraction. When Fr. A was purified and fractionated by DEAE-cellulose and Sepbadex G-200 gel filtration chromatographies into four fractions which were designated B, C, C-I and C-II. The tumor inhibition ratios of these fractions ranged from 46% to 75% against the solid forms of sarcoma 180 in ICR mice at doses of 20 and 50 mg/kg/day when given intraperitoneally. Especially, Fr. C which was named Collyban(CB) exhibited a marked life-prolonging effect of the mice against ascitic forms of sarcoma 180 at a dose of 50 mg via i.p. administration. To extend spectra of the antitumor activities and eliminate the effects of allograft rejection, the characterization of antitumor effects of CB was performed in syngeneic host-tumor systems. It did not show any antitumor activity against L1210 murine leukemia in $CD_2Fl$ mice but prolonged their life span against ascitic forms of $MM_{46}$ carcinoma in $C_3H/He$ mice. Also it exhibited antitumor activity against human cervical cancer HeLa cells that were xenografted into nude mice having BALB/c genetic backgrounds by the i.p. injection at a dose of 100 mg/kg/day. In order to characterize the antitumor components, CB was examined by chemical analysis. It was acidic protein-bound polysaccharides composed of 31% polysaccharide, 27% protein and 3% hexosamine. CB was fractionated into two fractions, Fr. C-I(M.W.: 500 Kd) and Fr. C-II(M.W.:30 and 8 Kd) by Sephadex G-200 gel filtration chromatography.