Isolation, Culture, Cryopreservation, and Preparation of Umbilical Cord-Derived Mesenchymal Stem Cells as a Final Cellul
Mesenchymal stem cells have gained popularity in cell-based therapies due to their regenerative capabilities, immunomodulation properties, and paracrine activity through trophic factors. It is of utmost importance to establish clinical-grade procedures fo
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Isolation, Culture, Cryopreservation, and Preparation of Umbilical Cord-Derived Mesenchymal Stem Cells as a Final Cellular Product Under Good Manufacturing Practices–Compliant Conditions Nurullah Aydog˘du, Olga Nehir O¨ztel, and Erdal Karao¨z Abstract Mesenchymal stem cells have gained popularity in cell-based therapies due to their regenerative capabilities, immunomodulation properties, and paracrine activity through trophic factors. It is of utmost importance to establish clinical-grade procedures for the preparation of the mesenchymal stem cells for clinical applications. Here, we describe detailed procedures for isolation, culture, cryopreservation, and preparation of mesenchymal stem cells derived from umbilical cord as a final product under good manufacturing practices–compliant conditions. Key words Umbilical cord, Stem cells, Cell isolation, Culture, Cell therapy, GMP
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Introduction Stem cell-based therapies have emerged as a novel approach in the field of regenerative medicine for the treatment of various diseases in order to repair and/or replace the impaired cells and tissues [1]. Mesenchymal stem cells (MSCs) are well-characterized, selfrenewable, multipotent adult stem cells that possess the ability to differentiate into various cell types of mesodermal origin [2]. MSCs can be isolated from different tissues, and can easily be expanded ex vivo and converted to mesodermal cell lineages [3]. One of the most attractive features of MSCs for clinical applications is their immunomodulation properties. They act as immune-conductors by producing and releasing various types of trophic factors in order to stimulate the neighboring parenchymal cells to start repairing the damaged tissue [4, 5]. These trophic factors possess different functions such as the enhancement of angiogenesis, stimulation of proliferation and differentiation, and modulation of the immune system [6]. Stem cell–based therapy is part of a broad term advanced therapy medicinal products (ATMPs) that refer to the clinical use
Nurullah Aydog˘du et al.
of medicinal products based on genes, cells, and tissues [7, 8]. The clinical use of ATMPs is regulated by the European Regulation EC 1394/2007 and the production of ATMPs must comply with good manufacturing practices (GMPs) which ensures the quality and purity of the final product through the control of all manufacturing processes and facilities [9]. GMP regulations include quality control assessments that are performed during the production as well as before the release of the cells for clinical use in order to assure the purity, quality, and safety of the cellular product. These assessments include cell number and cell viability assessment, characterization of cells, expression analysis of stemness genes, mycoplasma detection, analysis of telomerase activity, microbiological control, and bacterial endotoxin analysis. All methods for these analyses are validated and standard operating procedures (SOPs) are established based on these validation procedures [10, 11]. Here, we describe in detail t
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