Primary 3D Culture of Human Thyroid Follicle-Like Structures in Platelet Lysate-Based Gel
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Cell Technologies in Biology and Medicine, No. 2, August, 2020
Primary 3D Culture of Human Thyroid Follicle-Like Structures in Platelet Lysate-Based Gel
N. S. Sergeeva1,2, V. A. Kirsanova1, Yu. D. Khesuani3, I. K. Sviridova1, T. E. Skachkova1, V. A. Mironov3, A. P. Polyakov1, and A. D. Kaprin1 Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 2, pp. 104-111, June, 2020 Original article submitted November 16, 2018 The results of 3D culturing of human thyroid follicle-like structures in a gel based on platelet lysate at the gel-air interface are presented. During culturing up to 4 months, no new follicle-like structures were formed and none were destroyed. During the first 2 months, most follicle-like structures increased in size; then, their grown decelerated, but they retained viability. Ki-67+ cells were observed in the majority of follicle-like structures. Most of them produced thyroglobulin. Follicle-like structures get closer, the number of contacts between them increased, and cluster appeared. Thus, the developed 3D culturing system in a gel based on platelet lysate is an adequate approach for maintaining structure and functional activity of human follicle-like structures in vitro for at least 2 months. Key Words: thyroid gland; primary cultures; follicle-like structures; platelet lysate; gel
Culturing and development of technology for thyroid follicle (TF) expansion in vitro is in the focus of many laboratories over the past 30 years. This is due to the absence of adequate models for studying the regulation of physiological processes in the normal thyroid gland (TG) and the mechanisms of pathology development. Moreover, the ability to scale up the number of TF is the first step towards creating tissue-engineered constructs of TG for recovery of its anatomy and function after surgical interventions. Several research groups observed the formation of follicle-like structures (FS) in 2D-cultures of thyrocytes [5], but they quickly degraded and the cells formed a monolayer. Several media and cultures have been tested in attempts to stabilize FS [1,9,11,16]. However, these FS were often inverted and failed to form an adequate basal membrane [4,10,16]. Moscow P. A. Hertsen Research Oncological Institute — Affiliated Branch of National Medical Research Center of Radiology, Ministry of Health of the Russian Federation; 2N. I. Pirogov Russian National Research Medical University, Ministry of Health of the Russian Federation, Moscow, Russia; 33D Bioprinting Solutions Company, Moscow, Russia. Address for correspondence: [email protected]. N. S. Sergeeva 1
Some progress, for example, FS conversion, was achieved by transferring them to a 3D-collagen gel [4] and culturing on the gel-air interface [13]. The possibilities of formation of new FS in this system [1,6,7] and culturing native TF from TG tissue and containing (unlike newly formed TF) not only TC, but also C cells [12-15] were demonstrated. However, long-term maintenance of functionality of these TF remains an open question. We have develo
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