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**Chapter 3**

**Provisional chapter**

**"Cell-Free Therapeutics" from Components Secreted by**

Regenerative medicine is a fast growing multidisciplinary field aiming at the regeneration or replacement of damaged cells, tissues, or organs. Adult multipotent mesenchymal stromal cells (MSCs) are often used as a principal therapeutic tool in this field. Along with differentiation potency, MSCs secrete a wide spectrum of paracrine factors and extracellular vesicles participating in tissue repair and regeneration. Thus, for injuries that require trophic stimulation, cell survival support, and/or resident stem cells activation to be restored, one can apply MSC-conditioned medium, a combination of products and extracellular vesicles in cell culture growth medium, secreted by MSC. It could mediate most of beneficial regenerative effects of MSC without possible side effects of using MSC themselves. However, before the clinical application of this promising biopharmaceutical, several issues such as manufacturing protocols, quality control, and others must be addressed. Subsequently, we highlight the questions considering donor material variability, manufacturing, cell culture

medium and auxiliary components selection, and potency tests development.

medium, quality control, regenerative medicine

**Keywords:** biopharmaceuticals, multipotent mesenchymal stromal cells, conditioned

Regenerative medicine is a fast growing multidisciplinary field aiming at the regeneration or replacement of damaged cells, tissues, or organs. Adult stem and progenitor cells represent

**"Cell-Free Therapeutics" from Components Secreted** 

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,

distribution, and reproduction in any medium, provided the original work is properly cited.

DOI: 10.5772/intechopen.78605

**Mesenchymal Stromal Cells as a Novel Class of**

**by Mesenchymal Stromal Cells as a Novel Class of** 

**Biopharmaceuticals**

**Biopharmaceuticals**

Anastasia Yu. Efimenko

Anastasia Yu. Efimenko

**Abstract**

**1. Introduction**

Georgy D. Sagaradze, Peter P. Nimiritsky, Zhanna A. Akopyan, Pavel I. Makarevich and

Georgy D. Sagaradze, Peter P. Nimiritsky, Zhanna A. Akopyan, Pavel I. Makarevich and

Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.78605

#### **"Cell-Free Therapeutics" from Components Secreted by Mesenchymal Stromal Cells as a Novel Class of Biopharmaceuticals "Cell-Free Therapeutics" from Components Secreted by Mesenchymal Stromal Cells as a Novel Class of Biopharmaceuticals**

DOI: 10.5772/intechopen.78605

Georgy D. Sagaradze, Peter P. Nimiritsky, Zhanna A. Akopyan, Pavel I. Makarevich and Anastasia Yu. Efimenko Georgy D. Sagaradze, Peter P. Nimiritsky, Zhanna A. Akopyan, Pavel I. Makarevich and Anastasia Yu. Efimenko

Additional information is available at the end of the chapter Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.78605

#### **Abstract**

Regenerative medicine is a fast growing multidisciplinary field aiming at the regeneration or replacement of damaged cells, tissues, or organs. Adult multipotent mesenchymal stromal cells (MSCs) are often used as a principal therapeutic tool in this field. Along with differentiation potency, MSCs secrete a wide spectrum of paracrine factors and extracellular vesicles participating in tissue repair and regeneration. Thus, for injuries that require trophic stimulation, cell survival support, and/or resident stem cells activation to be restored, one can apply MSC-conditioned medium, a combination of products and extracellular vesicles in cell culture growth medium, secreted by MSC. It could mediate most of beneficial regenerative effects of MSC without possible side effects of using MSC themselves. However, before the clinical application of this promising biopharmaceutical, several issues such as manufacturing protocols, quality control, and others must be addressed. Subsequently, we highlight the questions considering donor material variability, manufacturing, cell culture medium and auxiliary components selection, and potency tests development.

**Keywords:** biopharmaceuticals, multipotent mesenchymal stromal cells, conditioned medium, quality control, regenerative medicine

#### **1. Introduction**

Regenerative medicine is a fast growing multidisciplinary field aiming at the regeneration or replacement of damaged cells, tissues, or organs. Adult stem and progenitor cells represent

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

a pivotal tool in regenerative medicine. It is important to highlight mesenchymal stromal cells (MSCs) as the most popular source for cell therapy and tissue engineering. However, a large amount of experimental data indicate that MSC effects on regeneration are mostly mediated by their ability to produce a wide range of bioactive molecules, and the use of MSCconditioned medium (CM), a complex of the factors secreted to cell culture growth medium, as a distinct biopharmaceutical drug can be a rational alternative to direct MSC therapy. MSC CM could be considered as "сell-free therapeutics" since this product is devoid of MSC themselves in its final formulation, yet possess significant therapeutic potency. It contains components secreted by MSC and reproduced the effects of MSC-based cell therapy.

cluster in MSC secretome is composed of extracellular matrix (ECM) proteins [6]. Such protein profile is in line with the stromal characteristics of adipose MSC. Adequate production of ECM components is necessary for tissue homeostasis and regeneration, because these molecules not only provide a scaffold for cells and soluble molecules but also regulate angiogenesis, neurogenesis, and inflammation. In addition, a large amount of data indicates that apart from soluble factors MSCs secrete regulatory non-coding RNA (e.g., micro RNA) within extracellular vesicles (EVs). The release of these small RNA by MSC can play a role in stem cell niche maintenance by controlling and tuning proliferation, differentiation, and homing. Particularly, microRNA regulates diverse biological processes, including growth and differ-

"Cell-Free Therapeutics" from Components Secreted by Mesenchymal Stromal Cells as a Novel…

http://dx.doi.org/10.5772/intechopen.78605

49

The function of adult stem cells includes the local or remote replacement of senescent or damaged cells along with maintaining their own pool. Stem cells supported by other niche components can participate in the repair of small lesions of a skin, liver, intestines, kidney, and bone marrow. However, stem cells could not cope with more serious injuries without more substantial support [9]. MSCs are important for maintaining the niche of stem cells; therefore, they can participate in stem cell potentiation to respond to damage, stimulate the survival of stem cells, and, thereby, maintain the structural and functional integrity of the niche. For example, in such serious damage as myocardium ischemia/reperfusion injury, MSC mediated its cardioprotective paracrine effect by secreting exosomes which reduced infarct size in a mouse model [10]. As in majority of animal models and clinical studies, only limited or no engraftment at all was often observed, one should consider paracrine MSC function as principal effector for tissue regeneration after, at least, systemic MSC injection for different

**3. Rationale for use of MSC and their cell-free derivatives in** 

MSCs are the most commonly used cells for cell-based therapy as they do not form teratomas, confer low immunogenicity, and are free of strict ethical concerns [12]. Despite these reasons, the difficulty to trace cell fate and survival in recipient has been a significant obstacle for understanding the mechanisms of the clinical efficacy that can be variable [13]. Furthermore, MSCs are highly heterogeneous and cannot be fully characterized in the context of identity and, finally, potency. At the same time, MSC CM was shown to mediate most of MSC beneficial effects. MSC CM includes various components such as cytokines, growth factors, ECM proteins and factors involved in ECM remodeling, different types of EV, and others. Using cell-free products based on biologically active factors secreted by stem and progenitor cells allows to significantly reduce the risks associated with a direct cell injection, while maintaining efficacy under wide manufacturing scalability and modification potential like fractionation, concentration, and combination with various carriers [14]. Therefore, the application of "cell-free therapeutics" based on the components secreted by MSC as a novel class of biopharmaceuticals represents a rapidly developing and promising approach in

entiation of stem cells [7, 8].

injuries [11].

**regenerative medicine**

regenerative medicine.

"Cell-free therapeutics" have various advantages in overcoming the limitations and risks associated with the cell-based therapy. Despite the outstanding preclinical and clinical efficacy of MSC CM, there is still no generally accepted regulating approaches for CM standardization and quality control. This chapter reviews the current state of art in the development of MSC CM-based medicinal products and describes the crucial issues concerning the production and quality control of this promising class of biopharmaceuticals.
