Genes & CellsGenes & Cells2313-18292500-2562Human Stem Cells Institute139206Research ArticleIsolation and culturing of chondrocytes from different sourcesКymI. I.redaktor@celltranspl.ru151220061448512601202326012023Copyright © 2006, Eco-Vector2006<p>The investigation was aimed to establish conditions for isolation and culturing cells of cartilaginous tissue procured from different sources.</p>
<p>Methods of chondrocytes obtaining and cultur<span style="text-decoration: line-through;">e</span>ing were compared. The chondrocytes were procured from human fetal cartilaginous tissue and the spinal column of patients with idiopathic scoliosis, as well as intervertebral disks of a new-born dog backbone.</p>
<p>The investigations of obtaining and culturing of chondrocytes from different sources showed that using collagenase solutions during 12-16 h at 37С with continuous interfusion allowed to receive cells suspension with more than 90% viability. The maximal amount of viable chondrocytes was derived while segregating cells using 0.15% collagenase. Нuman fetal cartilaginous tissue and that of dog intervertebral disks is being dissociated in collagenase for 5-8 h, tissue from the spinal column of patients with idiopathic scoliosis for 18-20 h.</p>
<p>While culturing human fetal chondrocytes had the best proliferating capability. The culture of chondrocytes from the spinal column of idiopathic scoliosis patients had low proliferation index and a short culturing time as compared with fetal culture.</p>chondrocytescell cultureculturingcollagenasetripsinhyaluronidaseхондроцитыкультура клетоккультивированиеколлагеназатрипсингиалуронидаза[Oakes B.W. Orthopaedic tissue engineering: from laboratory to the clinic. Bone and join disorders: prevention and control. The Medical Journal of Australia. Bone and Joint Disorders: Prevention and Control. 2004; 180: 35-8.][Vacanti J. et al. Tissue engineering in orthopedic surgery. Orthopedic Clinics of North America 2000; 31(3): 351-6.][Волков А.В. Тканевая инженерия в ортопедии: из лаборатории в клинику (мини-обзор). http://celltranspl.ru/journal/publications/?MESSAGES[1]=SHOW_PUBLICATION&PUBLICATION_ID=596.][Применение клеточных технологий в медицине. Центр Иммунотерапии и Клеточных Технологий. http://www.transplantation.ru/ru/stem-cell-therapy.php.][Гольдберг Е.Д., Дыгай А.М., Шахов В.П. Методы культуры ткани в гематологии. Томск: Изд-во Томского ун-та; 1992.][Фрешни Р. Культура животных клеток. Методы. М.: Мир; 1989: 117-33.][Пинаев Г.П. Методы культивирования клеток: Сборник научных трудов. Л.: Наука; 1988.][Богрянцева М.П., Трошкова Г.П., Мартынец Л.Д. и др. Некоторые аспекты производства и применения трипсина сухого стерильного для культур клеток. Международный конгресс «Биотехнология: состояние и перспективы развития». Москва, 14-18 октября, 2002 г. М.: ЗАО «ПИК «Максима»», РХТУ им. Д.И. Менделеева 2002: 98.][Jakob M., Demarteau O., Schafer D. et al. Enzymatic digestion of adult human articular cartilage yields a small fraction of the total available cells. Connect. Tissue Res. 2003; 44(3-4): 173-80.][Barbero A., Grogan S., Schafer D. et al. Age related changes in human articular chondrocyte yield, proliferation and post-expansion chondrogenic capacity. OsteoArthritis and Cartilage 2004; 12: 476Œ84.][Héraud F., Héraud A., Harmand M.-F. Apoptosis in normal and osteoarthritic human articular cartilage. Ann. Rheum. Dis. 2000; 59: 959-65.][Cornelissen M., Verbruggen G., Malfait A.M. et al. The study of representative populations of native aggrecan aggregates synthesized by human chondrocytes in vitro. J. Tiss. Cult. Meth. 1993; 15: 139Œ46.][Dharmavaram R.M., Liu G., Mowers S.D., Jimenez S.A. Detection and Characterization of Sp1 Binding Activity in Human Chondrocytes and Its Alterations during Chondrocyte Dedifferentiation. J. boil. chem. 1997; 272(43): 26918-25.][Зубов Д.А., Попандопуло А.Г., Слипченко И.О. и др. Морфологические особенности постнатальных хондроцитов человека in vitro. Цитология 2004; 46(10): 917.][Зайдман А.М. Идиопатический сколиоз: морфология, биохимия, генетика. Новосибирск: Новосиб. ун-та; 1993.]