Gaps in the knowledge of human platelet lysate as a cell culture supplement for cell therapy: a joint publication from the AABB and the International Society for Cell &amp; Gene Therapy

Bieback, Karen; Fernández‐Muñoz, Beatriz; Pati, Shibani; Schäfer, Richard

doi:10.1111/trf.15483

Cited by 60 publications

(25 citation statements)

References 106 publications

(206 reference statements)

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“…These assays must be quick, easy and should not require trained personnel if they are to be used to release each cell batch in a clinical setting and if they are to fit with operation theatre logistics (thawing, testing and infusing within a couple of hours). In theory, potency assays could be therapy-specific and must indicate cell functionality; in other words, “mechanism(s) of action” [70]. How much these assays correlate with the in vivo niche is also of great importance.…”

Section: Discussionmentioning

confidence: 99%

The impact of cryopreservation on bone marrow-derived mesenchymal stem cells: a systematic review

2019

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Mesenchymal stem cells (MSCs) represent an invaluable asset for the field of cell therapy. Human Bone marrow-derived MSCs (hBM-MSCs) are one of the most commonly used cell types in clinical trials. They are currently being studied and tested for the treatment of a wide range of diseases and conditions. The future availability of MSCs therapies to the public will require a robust and reliable delivery process. Cryopreservation represents the gold standard in cell storage and transportation, but its effect on BM-MSCs is still not well established. A systematic review was conducted to evaluate the impact of cryopreservation on BM-MSCs and to attempt to uncover the reasons behind some of the controversial results reported in the literature. Forty-one in vitro studies were analysed, and their results organised according to the cell attributes they assess. It was concluded that cryopreservation does not affect BM-MSCs morphology, surface marker expression, differentiation or proliferation potential. However, mixed results exist regarding the effect on colony forming ability and the effects on viability, attachment and migration, genomic stability and paracrine function are undefined mainly due to the huge variabilities governing the cryopreservation process as a whole and to the lack of standardised assays.

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Section: Discussionmentioning

confidence: 99%

The impact of cryopreservation on bone marrow-derived mesenchymal stem cells: a systematic review

2019

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“…A critical aspect in the clinical translation of MSC-based therapies is the use of safe and standardized culture conditions. Although commonly used for MSC expansion, several limitations of xenogeneic fetal bovine serum (FBS) supplementation have been highlighted, and current recommendations from health authorities advocate the use of 'xeno-free' protocols whenever possible (Bieback et al, 2019). Accordingly, xeno-free alternatives to FBS, such as human platelet lysate (HPL), have emerged (Shanbhag et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Xeno-Free Spheroids of Human Gingiva-Derived Progenitor Cells for Bone Tissue Engineering

Shanbhag

Suliman

Bolstad

et al. 2020

Front. Bioeng. Biotechnol.

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Gingiva has been identified as a minimally invasive source of multipotent progenitor cells (GPCs) for use in bone tissue engineering (BTE). To facilitate clinical translation, it is important to characterize GPCs in xeno-free cultures. Recent evidence indicates several advantages of three-dimensional (3D) spheroid cultures of mesenchymal stromal cells (MSCs) over conventional 2D monolayers. The present study aimed to characterize human GPCs in xeno-free 2D cultures, and to test their osteogenic potential in 3D cultures, in comparison to bone marrow MSCs (BMSCs). Primary GPCs and BMSCs were expanded in human platelet lysate (HPL) or fetal bovine serum (FBS) and characterized based on in vitro proliferation, immunophenotype and multilineage differentiation. Next, 3D spheroids of GPCs and BMSCs were formed via self-assembly and cultured in HPL. Expression of stemness-(SOX2, OCT4, NANOG) and osteogenesis-related markers (BMP2, RUNX2, OPN, OCN) was assessed at gene and protein levels in 3D and 2D cultures. The cytokine profile of 3D and 2D GPCs and BMSCs was assessed via a multiplex immunoassay. Monolayer GPCs in both HPL and FBS demonstrated a characteristic MSC-like immunophenotype and multi-lineage differentiation; osteogenic differentiation of GPCs was enhanced in HPL vs. FBS. CD271 + GPCs in HPL spontaneously acquired a neuronal phenotype and strongly expressed neuronal/glial markers. 3D spheroids of GPCs and BMSCs with high cell viability were formed in HPL media. Expression of stemness-and osteogenesisrelated genes was significantly upregulated in 3D vs. 2D GPCs/BMSCs; the latter was independent of osteogenic induction. Synthesis of SOX2, BMP2 and OCN was confirmed via immunostaining, and in vitro mineralization via Alizarin red staining. Finally, secretion of several growth factors and chemokines was enhanced in GPC/BMSC spheroids, while that of pro-inflammatory cytokines was reduced, compared to monolayers. In summary, monolayer GPCs expanded in HPL demonstrate enhanced osteogenic differentiation potential, comparable to that of BMSCs. Xeno-free spheroid culture further enhances stemness-and osteogenesis-related gene expression, and cytokine secretion in GPCs, comparable to that of BMSCs.

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“…6,7 In regenerative medicine, human platelet lysate (hPL), which is produced by freezing and thawing human platelets to release growth factors and trophogens in a lysate, is an effective growth factor supplement for several cell types including articular chrondrocytes, endothelial cells, dendritic cells, and osteoblasts. 3 hPL enhanced corneal endothelial cell proliferation and survival in vitro relative to FBS. 8 In clinical settings, platelet enriched plasma provides an important source of trophogens and growth factors facilitating stem-cellmediated tissue regeneration and repair.…”

Section: Introductionmentioning

confidence: 88%

“…Exemplifying its rich source of trophogens, serum from animal origin such as fetal bovine serum (FBS) is widely used in research applications and preclinical discovery. 3 However, cell culture with FBS in vitro does not mimic human microenvironments. This limits the translational applications of FBS, underscoring the need for effective substitutes.…”

Section: Introductionmentioning

confidence: 99%

Enhancing Chimeric Antigen Receptor T Cell Anti-tumor Function through Advanced Media Design

Ghassemi

Martinez-Becerra

Master

et al. 2020

Molecular Therapy - Methods & Clinical Development

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Effective chimeric antigen receptor (CAR)-T cell therapy is dependent on optimal cell culture methods conducive to the activation and expansion of T cells ex vivo , as well as infection with CAR. Media formulations used in CAR-T cell manufacturing have not been optimized for gene delivery, cell expansion, and overall potency. Bioactive components and derivatives that support the generation of functionally-competent T cell progeny with long-lasting persistence are largely undefined. Current media formulations rely on fetal bovine serum (FBS) or human serum (HS), which suffer from a lack of consistency or supply issues. We recognize that components of blood cellular fractions that are absent in serum may have therapeutic value. Here we investigate whether a concentrated growth factor extract, purified from human transfusion grade whole blood fractions, and marketed as PhysiologixTM xeno-free (XF) hGFC (Phx), supports CAR-T cell expansion and function. We show that Phx supports T cell proliferation in clinical and research-grade media. We also show that Phx treatment enhances lentiviral-mediated gene expression across a wide range of multiplicity of infections (MOIs). We compared the ability of anti-GD-2 CAR-T cells expanded ex vivo in medium conditioned with either Phx or HS to clear tumor burden in a human xenograft model of neuroblastoma. We show that T cells expanded in Phx have superior engraftment and potency in vivo , as well as CAR-induced cytolytic activity in vitro . Metabolomic profiling revealed several factors unique to Phx that may have relevance for CAR-T cell preclinical discovery, process development, and manufacturing. In particular, we show that carnosine, a biogenic amine modestly enriched in Phx relative to HS, enhances lentiviral gene delivery in activated T cells. By limiting extracellular acidification, carnosine enhances the metabolic fitness of T cells, shifting their metabolic profile from an acidic, stressed state toward an oxidative, energetic state. These findings are very informative regarding potential derivatives to include in medium customized for gene delivery and overall potency for T cell adoptive immunotherapies.

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Gaps in the knowledge of human platelet lysate as a cell culture supplement for cell therapy: a joint publication from the AABB and the International Society for Cell & Gene Therapy

Cited by 60 publications

References 106 publications

The impact of cryopreservation on bone marrow-derived mesenchymal stem cells: a systematic review

The impact of cryopreservation on bone marrow-derived mesenchymal stem cells: a systematic review

Xeno-Free Spheroids of Human Gingiva-Derived Progenitor Cells for Bone Tissue Engineering

Enhancing Chimeric Antigen Receptor T Cell Anti-tumor Function through Advanced Media Design

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