Manual cryopreservation of human alveolar periosteal tissue segments: Effects of pre-culture on recovery rate

被引:11
作者
Kawase, Tomoyuki [1 ]
Kogami, Hiroyuki [1 ]
Nagata, Masaki [2 ]
Uematsu, Kohya [2 ]
Okuda, Kazuhiro [3 ]
Burns, Douglas M. [4 ]
Yoshie, Hiromasa [3 ]
机构
[1] Niigata Univ, Inst Med & Dent, Dept Tissue Regenerat & Reconstitut, Div Oral Bioengn, Niigata 9518514, Japan
[2] Niigata Univ, Inst Med & Dent, Dept Oral Hlth Sci, Div Oral & Maxillofacial Surg, Niigata 9518514, Japan
[3] Niigata Univ, Inst Med & Dent, Dept Oral Biol Sci, Div Periodontol, Niigata 9518514, Japan
[4] Med Res Serv, Dept Vet Affairs Med Ctr, Kansas City, MO 64105 USA
关键词
Periosteum; Human; Cryopreservation; Dimethyl sulfoxide; Fetal bovine serum; Cell outgrowth; REPLACEMENT; VIABILITY; ALBUMIN; CELLS;
D O I
10.1016/j.cryobiol.2011.03.004
中图分类号
Q [生物科学];
学科分类号
090105 [作物生产系统与生态工程];
摘要
Cultured human periosteal sheets constitute a promising grafting material for periodontal tissue regenerative therapy. However, preparation of these sheets usually requires six weeks or longer, and this lengthy commitment and delay limits both clinical applicability and availability. The aim of this study is to develop an efficient, practical, cost-effective cryopreservation method for periosteal tissue segments (PTSs). Human PTSs were aseptically excised from alveolar bone and pre-cultured in Medium 199 + 10% fetal bovine serum (FBS) for the indicated number of days before they were slowly frozen down to -75 degrees C in a commercial freezing vessel using medium containing 10% dimethyl sulfoxide (Me2SO) and various concentrations of FBS. After fast-thawing at 37 degrees C, PTSs were again cultured, and their growth and responses to standard osteogenic induction were evaluated (vs. freshly excised PTSs). Proliferating cells were obtained at the highest levels from cryopreserved FTSs that were pre-cultured for 14 days before freezing. When a concentration of 50% or more FBS was included in the cryopreservation solution, cells migrated out more actively and grew faster. Importantly, osteoinduction up-regulated alkaline phosphatase (ALP) activity and osteoblastic marker mRNAs in cryopreserved PTS-derived sheets just as effectively as it did in native PTS-derived ones. These data suggest that pre-conditioned PTSs can be efficiently cryopreserved in a freezing solution containing high FBS by traditional manual cryopreservation methods without aid of a program freezer or more elaborate equipment. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码:202 / 209
页数:8
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