Growth and surface properties of dispase dissociated human fibroblasts |
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| Affiliation: | 1. Department of Biotechnology, Seoul Women''s University, 621 Hwarangro, Nowon-Gu, Seoul, Republic of Korea;2. Department of Biomedical Science, CHA University, 120 Haeryong-ro, Pocheon-shi, Gyeongghi-do, Republic of Korea;3. Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Republic of Korea;1. Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstr. 14, 35390 Giessen, Germany;2. Faculty of Biology and Chemistry, Justus Liebig University, Ludwigstr. 23, 35390 Giessen, Germany;3. Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Project Group Bioresources, Winchester Str. 2, 35394 Giessen, Germany;4. Department of Chemical Engineering, Kansas State University, 1005 Durland Hall, Manhattan, KS 66506, USA;1. SpheroTec GmbH, Am Klopferspitz 19, 82152 Martinsried, Germany;2. Department of General, Visceral, Transplantation, Vascular and Thoracic Surgery, University of Munich, Campus Großhadern, Marchioninistr. 15, 81377 Munich, Germany;1. Laboratório de Pesquisa em Patologia, Universidade Federal de Ciências da Saúde de Porto Alegre, Rua Sarmento Leite, 245, Porto Alegre - Rio Grande do Sul, Brazil;2. Laboratório de Pesquisa em Biologia Celular, Universidade Federal de Ciências da Saúde de Porto Alegre, Rua Sarmento Leite, 245, Porto Alegre - Rio Grande do Sul, Brazil;3. Laboratório de Bioquímica Celular, Departamento de Bioquímica, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600, Porto Alegre - Rio Grande do Sul, Brazil |
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| Abstract: | The dissociation of human fibroblast cultures with the bacterial neutral protease (Dispase II) was monitored by viability and growth measurements and was compared to the effect of trypsin and EDTA. Cell suspensions with high viability and excellent growth were obtained after 10 min incubation at 37 °C in 4 U/ml dispase in 0.02% EDTA. A two to three-fold increase in mitotic index occurred in the cultures within 48 h after dispase dissociation. The initial rate of aggregation was comparable to that of trypsin or EDTA dissociated cells, but attachment to a substratum and agglutination by Wheat Germ Agglutinin were markedly enhanced. The results indicate that dispase-EDTA provides a valuable alternative to the enzymatic dissociation with trypsin. Moreover, it is an additional tool for the dissociation of cultured cells and for the study of the surface properties of single cells. |
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