Toward a Clinical-Grade Expansion of Mesenchymal Stem Cells from Human Sources: A Microcarrier-Based Culture System Under Xeno-Free Conditions

Author(s): Santos FD, Andrade PZ, Abecasis MM, Gimble JM, Chase LG, et al.

Abstract

The immunomodulatory properties of mesenchymal stem cells (MSCs) make them attractive therapeutic agents for a wide range of diseases. However, the highly demanding cell doses used in MSC clinical trials (up to millions of cells/kg patient) currently require labor intensive methods and incur high reagent costs. Moreover, the use of xenogenic (xeno) serum-containing media represents a risk of contamination and raises safety concerns. Bioreactor systems in combination with novel xeno-free medium formulations represent a viable alternative to reproducibly achieve a safe and reliable MSC doses relevant for cell therapy. The main goal of the present study was to develop a complete xeno-free microcarrier-based culture system for the efficient expansion of human MSC from two different sources, human bone marrow (BM), and adipose tissue. After 14 days of culture in spinner flasks, BM MSC reached a maximum cell density of (2.0±0.2)×10⁵ cells·mL⁻¹ (18±1-fold increase), whereas adipose tissue-derived stem cells expanded to (1.4±0.5)×10⁵ cells·mL⁻¹ (14±7-fold increase). After the expansion, MSC expressed the characteristic markers CD73, CD90, and CD105, whereas negative for CD80 and human leukocyte antigen (HLA)-DR. Expanded cells maintained the ability to differentiate robustly into osteoblast, adipocyte, and chondroblast lineages upon directed differentiation. These results demonstrated the feasibility of expanding human MSC in a scalable microcarrier-based stirred culture system under xeno-free conditions and represent an important step forward for the implementation of a Good Manufacturing Practices-compliant large-scale production system of MSC for cellular therapy.

Similar Articles

Non-expanded adipose stromal vascular fraction cell therapy for multiple sclerosis

Author(s): Riordan NH, Ichim TE, Min WP, Wang H, Solano F, et al.

Isolation and characterization of multi-potent stem cells from human orbital fat tissues

Author(s): Ho JH, Ma WH, Tseng TC, Chen YF, Chen MH, et al.

Isolation and characterization of canine adipose-derived mesenchymal stem cells

Author(s): Neupane M, Chang CC, Kiupel M,Yuzbasiyan-Gurkan V

Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives

Author(s): Planat-Benard V, Silvestre JS, Cousin B, Andre M, Nibbelink M, et al.

Progenitor-enriched adipose tissue transplantation as rescue for breast implant complications

Author(s): Yoshimura K, Asano Y, Aoi N, Kurita M, Oshima Y, et al.

Autologous stem cells (adipose) and fibrin glue used to treat widespread traumatic calvarial defects: case report

Author(s): Lendeckel S, Jodicke A, Christophis P, Heidinger K, Wolff J, et al.

Adipose-derived stromal cells: Their identity and uses in clinical trials, an update

Author(s): Casteilla L, Planat-Benard V, Laharrague P,Cousin B

A simple modification of the separation method reduces heterogeneity of adipose-derived stem cells

Author(s): Griesche N, Luttmann W, Luttmann A, Stammermann T, Geiger H, et al.

Characterization of adipose tissue-derived cells isolated with the Celution system

Author(s): Lin K, Matsubara Y, Masuda Y, Togashi K, Ohno T, et al.

Sera of overweight people promote in vitro adipocyte differentiation of bone marrow stromal cells

Author(s): Di Bernardo G, Messina G, Capasso S, Del Gaudio S, Cipollaro M, et al.

Comparison of ex vivo expansion culture conditions of mesenchymal stem cells for human cell therapy

Author(s): Perez-Ilzarbe M, Diez-Campelo M, Aranda P, Tabera S, Lopez T, et al.

Inhibition of mesenchymal stromal cells by pre-activated lymphocytes and their culture media

Author(s): Valencic E, Loganes C, Cesana S, Piscianz E, Gaipa G, et al.

Ex vivo expansion of human mesenchymal stem cells in defined serum-free media

Author(s): Jung S, Panchalingam KM, Rosenberg L,Behie LA

Optimizing proliferation and characterization of multipotent stem cells from porcine adipose tissue

Author(s): Wang KH, Kao AP, Wangchen H, Wang FY, Chang CH, et al.

Intravenous administration of auto serum-expanded autologous mesenchymal stem cells in stroke

Author(s): Honmou O, Houkin K, Matsunaga T, Niitsu Y, Ishiai S, et al.

Expectations and strategies regarding islet transplantation: metabolic data from the GRAGIL 2 trial

Author(s): Badet L, Benhamou PY, Wojtusciszyn A, Baertschiger R, Milliat-Guittard L, et al.

Human CD34/CD90 ASCs are capable of growing as sphere clusters, producing high levels of VEGF and forming capillaries

Author(s): De Francesco F, Tirino V, Desiderio V, Ferraro G, D'Andrea F, et al.