Tissue Source and Cell Expansion Condition Influence Phenotypic Changes of Adipose-Derived Stem Cells

Mangum, Lauren H.; Natesan, Shanmugasundaram; Stone, Randolph; Wrice, Nicole L.; Larson, David A.; Florell, K; Christy, Barbara A.; Herzig, Maryanne C.; Andrew, P.; Christy, Robert J.

doi:10.1155/2017/7108458

Cited by 12 publications

(13 citation statements)

References 67 publications

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“…The procoagulant effects of CCTs in trauma patients must be considered as a safety release criterion for their clinical use. Consideration should be given to choosing cell sources expressing less TF and cell expansion methods that minimize TF expression should also be explored …”

Section: Discussionmentioning

confidence: 99%

Procoagulant in vitro effects of clinical cellular therapeutics in a severely injured trauma population

George

Prabhakara

Toledano-Furman

et al. 2020

Stem Cells Translational Medicine

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Clinical trials in trauma populations are exploring the use of clinical cellular therapeutics (CCTs) like human mesenchymal stromal cells (MSC) and mononuclear cells (MNC). Recent studies demonstrate a procoagulant effect of these CCTs related to their expression of tissue factor (TF). We sought to examine this relationship in blood from severely injured trauma patients and identify methods to reverse this procoagulant effect. Human MSCs from bone marrow, adipose, and amniotic tissues and freshly isolated bone marrow MNC samples were tested. TF expression and phenotype were quantified using flow cytometry. CCTs were mixed individually with trauma patients' whole blood, assayed with thromboelastography (TEG), and compared with healthy subjects mixed with the same cell sources. Heparin was added to samples at increasing concentrations until TEG parameters normalized. Clotting time or R time in TEG decreased relative to the TF expression of the CCT treatment in a logarithmic fashion for trauma patients and healthy subjects. Nonlinear regression curves were significantly different with healthy subjects demonstrating greater relative decreases in TEG clotting time. In vitro coadministration of heparin normalized the procoagulant effect and required dose escalation based on TF expression. TF expression in human MSC and MNC has a procoagulant effect in blood from trauma patients and healthy subjects. The procoagulant effect is lower in trauma patients possibly because their clotting time is already accelerated. The procoagulant effect due to MSC/MNC TF expression could be useful in the bleeding trauma patient; however, it may emerge as a safety release criterion due to thrombotic risk. The TF procoagulant effect is reversible with heparin.

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Section: Discussionmentioning

confidence: 99%

Procoagulant in vitro effects of clinical cellular therapeutics in a severely injured trauma population

George

Prabhakara

Toledano-Furman

et al. 2020

Stem Cells Translational Medicine

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“…In addition, the concentration of growth factors seems to be limited in both hS and FBS, and some authors found that the debris, that forms during PRP production, could slow down cell expansion [ 6 , 14 ]. However, the relative advantages of these different culture media additives are still widely debated and recent studies showed that particularly hPL could be a valid substitute for FBS, retaining stem cell phenotype (positive expression of CD (cluster of differentiation) CD73, CD90, CD105, but negative for CD34 and CD45) and multilineage differentiation capacity (osteogenic, adipogenic and chondrogenic) of hMSCs [ 15 – 17 ].…”

Section: Introductionmentioning

confidence: 99%

Human platelet lysate to substitute fetal bovine serum in hMSC expansion for translational applications: a systematic review

et al. 2020

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Background Foetal bovine serum (FBS), is the most commonly used culture medium additive for in vitro cultures, despite its undefined composition, its potential immunogenicity and possible prion/zoonotic transmission. For these reasons, significant efforts have been targeted at finding a substitute, such as serum free-media or human platelet-lysates (hPL). Our aim is to critically appraise the state-of-art for hPL in the published literature, comparing its impact with FBS. Materials and methods In June 2019 a systematic search of the entire Web of Science, Medline and PubMed database was performed with the following search terms: (mesenchymal stem cells) AND (fetal bovine serum OR fetal bovine calf) AND (human platelet lysate). Excluded from this search were review articles that were published before 2005, manuscripts in which mesenchymal stem cells (MSCs) were not from human sources, and when the FBS controls were missing. Results Based on our search algorithm, 56 papers were selected. A review of these papers indicated that hMSCs cultured with hPL showed a spindle-shaped elongated morphology, had higher proliferation indexes, similar cluster of differentiation (CD) markers and no significant variation in differentiation lineage (osteocyte, adipocyte, and chondrocyte) compared to those cultured with FBS. Main sources of primary hMSCs were either fat tissue or bone marrow; in a few studies cells isolated from alternative sources showed no relevant difference in their response. Conclusion Despite the difference in medium choice and a lack of standardization of hPL manufacturing, the majority of publications support that hPL was at least as effective as FBS in promoting adhesion, survival and proliferation of hMSCs. We conclude that hPL should be considered a viable alternative to FBS in hMSCs culture—especially with a view for their clinical use.

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“…Given that Stemulate™ is used to expand a range of cell types, including adipose-derived mesenchymal stem cells (MSCs) and bone-marrow MSCs, it could be an alternative supplement to increase chondrocyte proliferation for cartilage cell therapy (Badowski et al, 2017;Juhl et al, 2016;Mangum et al, 2017;Riis et al, 2016;Søndergaard et al, 2017). Stemulate™ could reduce time in monolayer culture and reduce exposure to tissue-culture plastic, which in turn could reduce the risk of de-differentiation.…”

Section: Introductionmentioning

confidence: 99%

Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

Sykes¹,

Kuiper²,

Richardson³

et al. 2018

eCM

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High hopes have been pinned on regenerative medicine strategies in order to prevent the progression of cartilage damage to osteoarthritis, particularly by autologous chondrocyte implantation (ACI). The loss of chondrocyte phenotype during in vitro monolayer expansion, a necessary step to obtain sufficient cell numbers, may be a key limitation in ACI. In this study, it was determined whether a shorter monolayer expansion approach could improve chondrogenic differentiation. The effects of two supplement types, foetal bovine serum (FBS) and Stemulate™ (a commercial source of human platelet lysate), on the expansion and re-differentiation potential of human chondrocytes, isolated from five individuals, were compared. Chondrocytes were expanded with 10 % FBS or 10 % Stemulate™. Pellets were cultured for 28 d in chondrogenic differentiation medium and assessed for the presence of cartilage matrix molecules and genes associated with chondrogenicity. Stemulate™ significantly enhanced the proliferation rate [average population doubling times: FBS, 25.07 ± 6.98 d (standard error of the mean, SEM) vs. Stemulate™, 13.10 ± 2.57 d (SEM)]. Sulphated glycosaminoglycans (sGAG), total collagen and qRT-PCR analyses of cartilage genes showed that FBS-expanded chondrocytes demonstrated significantly better chondrogenic capacity than Stemulate™-expanded chondrocytes. Histologically, FBS-expanded chondrocyte pellets appeared to be more stable, with a more intense staining for toluidine blue, indicating a greater chondrogenic capacity. Although Stemulate™ positively influenced chondrocyte proliferation, it had a negative effect on chondrogenic differentiation potential. This suggested that, in the treatment of cartilage defects, Stemulate™ might not be the ideal supplement for expanding chondrocytes (which maintained a chondrocyte phenotype) and, hence, for cell therapies (including ACI).

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Tissue Source and Cell Expansion Condition Influence Phenotypic Changes of Adipose-Derived Stem Cells

Cited by 12 publications

References 67 publications

Procoagulant in vitro effects of clinical cellular therapeutics in a severely injured trauma population

Procoagulant in vitro effects of clinical cellular therapeutics in a severely injured trauma population

Human platelet lysate to substitute fetal bovine serum in hMSC expansion for translational applications: a systematic review

Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

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