A Good Manufacturing Practice–grade standard protocol for exclusively human mesenchymal stromal cell–derived extracellular vesicles

Pachler, Karin; Lener, Thomas; Streif, Doris; Dunai, Zsuzsanna A.; Desgeorges, Alexandre; Feichtner, Martina; Öller, Michaela; Schallmoser, Katharina; Rohde, Eva; Gimona, Mario

doi:10.1016/j.jcyt.2017.01.001

Cited by 171 publications

(129 citation statements)

References 36 publications

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“…However, ALP activity in the H1 group remains beneath F10 and H10 groups at any time but D1. Interestingly, most studies determining hPL concentration in the context of osteogenic differentiation only determined calcium content [16,22,23,[25][26][27]29,33,53]; only a few studies assessed ALP activity [18,21] and, therefore, did not detect the described discrepancy in ALP activity and calcium deposition. Only 10% hPL-supplemented ODM showed a constantly better or equal osteogenic differentiation compared to FCS.…”

Section: Discussionmentioning

confidence: 99%

“…However, it remains unclear how hPL as a part of an osteogenic differentiation medium (ODM) influences the kinetics of the osteogenic differentiation of BMSC. Furthermore, the necessary concentrations of hPL reported differ from 2% to 10% [18,20,25,26,[29][30][31][32][33][34], pointing out that there is no established standard protocol for hPL supplementation in ODM for BMSC yet, although the concentration of serum supplement in media resulting in different concentrations of growth factors and other components of the serum influences cells in culture [1].…”

Section: Introductionmentioning

confidence: 99%

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Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Karadjian

Senger

Essers

et al. 2020

Cells

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Fetal calf serum (FCS) is frequently used as a growth factor and protein source in bone-marrow-derived mesenchymal stromal cell (BMSC) culture media, although it is a xenogenic product presenting multiple disadvantages including but not limited to ethical concerns. A promising alternative for FCS is human platelet lysate (hPL), which is produced out of human platelet concentrates and happens to be a stable and reliable protein source. In this study, we investigated the influence of hPL in an expansion medium (ESM) and an osteogenic differentiation medium (ODM) on the proliferation and osteogenic differentiation capacity of human BMSC. Therefore, we assessed population doublings during cell expansion, performed alizarin red staining to evaluate the calcium content in the extracellular matrix and determined the activity of alkaline phosphatase (ALP) as osteogenic differentiation correlates. The proliferation rate of BMSC cultured in ESM supplemented with hPL exceeded the proliferation rate of BMSC cultured in the presence of FCS. Furthermore, the calcium content and ALP activity was significantly higher in samples incubated in hPL-supplemented ODM, especially in the early phases of differentiation. Our results show that hPL can replace FCS as a protein supplier in cell culture media and does not negatively affect the osteogenic differentiation capacity of BMSC.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Karadjian

Senger

Essers

et al. 2020

Cells

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“…In early 2002, Lamparski et al (2002) described a production, purification, and characterization method for GMP-grade EVs from antigen presenting cells as a viable vaccine for cancer. Recently, Pachler et al (2017) developed a GMP standard protocol for human MEVs. More recently, a GMP-grade method for the large-scale preparation of EVs from human cardiac progenitor cells was described (Andriolo et al, 2018).…”

Section: Insufficient Clinical Grade Productionmentioning

confidence: 99%

Prospects and challenges of extracellular vesicle-based drug delivery system: considering cell source

et al. 2020

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Extracellular vesicles (EVs), including exosomes, microvesicles, and apoptotic bodies, are nanosized membrane vesicles derived from most cell types. Carrying diverse biomolecules from their parent cells, EVs are important mediators of intercellular communication and thus play significant roles in physiological and pathological processes. Owing to their natural biogenesis process, EVs are generated with high biocompatibility, enhanced stability, and limited immunogenicity, which provide multiple advantages as drug delivery systems (DDSs) over traditional synthetic delivery vehicles. EVs have been reported to be used for the delivery of siRNAs, miRNAs, protein, small molecule drugs, nanoparticles, and CRISPR/Cas9 in the treatment of various diseases. As a natural drug delivery vectors, EVs can penetrate into the tissues and be bioengineered to enhance the targetability. Although EVs' characteristics make them ideal for drug delivery, EV-based drug delivery remains challenging, due to lack of standardized isolation and purification methods, limited drug loading efficiency, and insufficient clinical grade production. In this review, we summarized the current knowledge on the application of EVs as DDS from the perspective of different cell origin and weighted the advantages and bottlenecks of EV-based DDS. ARTICLE HISTORY

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“…MSCs have been infused in patients with various disorders without any significant side effects (19). Additionally, bone marrow-derived MSCs have emerged as an attractive cellular source for the generation of clinical-grade exosomes for human therapies (20,21). Here, we report on the production process and the potential of MSCs in the generation and engineering Exosomes are extracellular vesicles produced by all cells with a remarkable ability to efficiently transfer genetic material, including exogenously loaded siRNA, to cancer cells.…”

Section: Introductionmentioning

confidence: 99%

Generation and testing of clinical-grade exosomes for pancreatic cancer

et al. 2018

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A Good Manufacturing Practice–grade standard protocol for exclusively human mesenchymal stromal cell–derived extracellular vesicles

Cited by 171 publications

References 36 publications

Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Prospects and challenges of extracellular vesicle-based drug delivery system: considering cell source

Generation and testing of clinical-grade exosomes for pancreatic cancer

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