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

Pérez-Ilzarbe, Maitane; Díez-Campelo, María; Aranda, Pablo; Tabera, Soraya; López, Tania; Cañizo, Consuelo del; Merino, Juana; Moreno, Cristina; Andreu, Enrique J.; Prósper, Felipe; Pérez-Simón, Josè Antonio

doi:10.1111/j.1537-2995.2009.02226.x

Cited by 91 publications

(62 citation statements)

References 30 publications

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“…This effect, however, may not necessarily hold true in ‘humanized' culture systems. Similar to our own yet unpublished data, bFGF appears to support expansion of FBS-supplemented bone marrow MSC cultures, but fails to do so in HPL-supplemented systems [105]. Neutralization of PDGF-AB/BB, TGF-β1 and FGF significantly diminished the proliferative effect of HPL, with the strongest effects seen by neutralizing FGF alone or in combination with PDGF-BB [77].…”

Section: Platelet Lysatesupporting

confidence: 51%

Platelet Lysate as Replacement for Fetal Bovine Serum in Mesenchymal Stromal Cell Cultures

Bieback

2013

Transfus Med Hemother

180

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Mesenchymal stromal cells (MSC) emerged as highly attractive in cell-based regenerative medicine. Initially thought to provide cells capable of differentiation towards mesenchymal cell types (osteoblasts, chondrocytes, adipocytes etc.), by and by potent immunoregulatory and pro-regenerative activities have been discovered, broadening the field of potential applications from bone and cartilage regeneration to wound healing and treatment of autoimmune diseases. Due to the limited frequency in most tissue sources, ex vivo expansion of MSC is required compliant with good manufacturing practice (GMP) guidelines to yield clinically relevant cell doses. Though, still most manufacturing protocols use fetal bovine serum (FBS) as cell culture supplement to isolate and to expand MSC. However, the high lot-to-lot variability as well as risk of contamination and immunization call for xenogenic-free culture conditions. In terms of standardization, chemically defined media appear as the ultimate achievement. Since these media need to maintain all key cellular and therapy-relevant features of MSC, the development of chemically defined media is still - albeit highly investigated - only in its beginning. The current alternatives to FBS rely on human blood-derived components: plasma, serum, umbilical cord blood serum, and platelet derivatives like platelet lysate. Focusing on quality aspects, the latter will be addressed within this review.

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Section: Platelet Lysatesupporting

confidence: 51%

Platelet Lysate as Replacement for Fetal Bovine Serum in Mesenchymal Stromal Cell Cultures

Bieback

2013

Transfus Med Hemother

180

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“…Those serum batches are similar due to extensive analysis of the manufacturer, but they are not equivalent. A further cause of heterogeneity is the use of human platelet lysate (hPL) for expansion of MSCs [25,[32][33][34][35][36]. Expansion in hPL is attractive because it does not contain animal components [32,35,37], which allows cells cultured in hPL to be used in clinical settings [38].…”

mentioning

confidence: 99%

The Impact of Cell Source, Culture Methodology, Culture Location, and Individual Donors on Gene Expression Profiles of Bone Marrow-Derived and Adipose-Derived Stromal Cells

Torensma

Prins

Schrama

et al. 2013

Stem Cells and Development

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Bone marrow (BM) stromal cells (MSCs), also known as mesenchymal stem cells, display a high degree of heterogeneity. To shed light on the causes of this heterogeneity, MSCs were collected from either human BM (n = 5) or adipose tissue (AT) (n = 5), and expanded using 2 different culture methods: one based on fetal calf serum, and one based on human platelet lysate. After initial expansion, MSCs were frozen, and the vials were transported to 3 different laboratories and grown for 1 passage using the same brand of culture plastic, medium, and supplements. Subsequently, the cells were harvested and assayed for their gene expression profile using the Affymetrix exon microarray platform. Based on gene expression profiles, the most discriminative feature was the anatomical harvesting site, followed by culture methodology. Remarkably, genes in the WNT pathway were expressed at higher levels in BM-derived MSCs than in AT-derived MSCs. Although differences were found between laboratories, cell culture location only slightly affects heterogeneity. Furthermore, individual donors contributed marginally to the observed differences in transcriptomes. Finally, BM-derived MSCs displayed the highest level of similarity, irrespective their culture conditions, when compared to AT-derived cells.

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“…4 Several clinical trials have been designed in an attempt to explore their therapeutic potential in acute graft-versus-host disease (GVHD), 5,6 while the information available on the efficacy of MSCs in the chronic GVHD setting is far more limited 7 (www.clinicaltrials.gov). We have previously shown that the expansion of MSC using human serum is feasible, 8 and their immunomodulatory properties were preserved and comparable with MSC expanded using FCS. In order to prevent viral or prion contamination, we designed a phase I/II clinical trial in order to evaluate the potential benefit of the infusion of MSC expanded using human serum (HS) among patients diagnosed with either refractory acute or chronic graft-versus-host dis- …”

Section: Introductionmentioning

confidence: 99%

Mesenchymal stem cells expanded in vitro with human serum for the treatment of acute and chronic graft-versus-host disease: results of a phase I/II clinical trial

Pérez-Simón¹,

López-Villar²,

Andreu³

et al. 2011

Haematologica

157

105

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MSC expansion procedure, cell characterization and obtaining platelet lysate were performed according to standard procedures and are shown in the Online Supplementary Appendix. Patients' characteristics and MSC infusionOverall, 18 patients diagnosed with either acute (n=10) or chronic (n=8) GVHD refractory to prior treatment were included in the study once written informed consent had been obtained. Refractory GVHD was defined as progression or absence of response to last treatment. Patients' characteristics are summarized in Table 2. Eleven patients had received reduced intensity conditioning and 6 received myeloablative conditioning. Fourteen patients had received hematopoietic stem cells from an unrelated donor and 7 received them from an HLA-mismatched donor. Eight patients had received GVHD prophylaxis based on a calcineurin inhibitor plus methotrexate.Inclusion criteria were: patients who had undergone an allogeneic stem cell transplant and developed GVHD refractory to conventional treatment; adequate cardiac and pulmonary functions; aged between 18 and 65 years; signed informed consent from patient and donor. Exclusion criteria were: patients who did not fulfill all of the inclusion criteria; progression of the hematologic disease; active infection; women who were either pregnant or at risk of pregnancy. The study was conducted between February 2007 and December 2009.Patients received 1-2x10 6 MSCs/kg intravenously in a single dose. Eventually, when a partial response was obtained or in the case of relapse after achieving complete remission, patients could receive a second dose of MSCs at least two weeks after the first infusion. Patients who were receiving 6-methylprednisolone were kept on the same doses for at least seven days after MSC infusion and a taper of 10% every five days was planned later when there was a response. Other immunosuppressive drugs were managed according to the criteria of the attending physician. Response to therapy was measured according to previously reported criteria. [9][10][11] Patients were taken off the study if fewer than 1x10 6 MSCs/kg were obtained after eight weeks of expansion. All patients receiving at least one dose of MSCs were included in the safety and efficacy analysis.The treatment protocol was reviewed and approved by the local authorities and ethical committee of all participanting centers.Mesenchymal cells for graft vs. host disease treatment haematologica | 2011; 96 (7) 1073 Statistical analysisVariables of the expansion procedure were analyzed from the day of inclusion in the trial, i.e. the day when informed consent was signed. Mean and median values and their corresponding 95% confidence intervals (CIs) and ranges were calculated for each continuous variable. Student's two-sample t test and Pearson's X 2 test were used to compare continuous and qualitative variables. In those comparisons where the number of cases precluded the use of parametric tests, the Mann-Whitney test and Fisher's exact test were used. To analyze patient outcome after infusi...

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

Cited by 91 publications

References 30 publications

Platelet Lysate as Replacement for Fetal Bovine Serum in Mesenchymal Stromal Cell Cultures

Platelet Lysate as Replacement for Fetal Bovine Serum in Mesenchymal Stromal Cell Cultures

The Impact of Cell Source, Culture Methodology, Culture Location, and Individual Donors on Gene Expression Profiles of Bone Marrow-Derived and Adipose-Derived Stromal Cells

Mesenchymal stem cells expanded in vitro with human serum for the treatment of acute and chronic graft-versus-host disease: results of a phase I/II clinical trial

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