Heparin and Derivatives for Advanced Cell Therapies

Laner-Plamberger, Sandra; Oeller, Michaela; Rohde, Eva; Schallmoser, Katharina; Strunk, Dirk

doi:10.3390/ijms222112041

Cited by 10 publications

(13 citation statements)

References 179 publications

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“…More standardized study protocols and a dose-response relationship would be required to define hypothetic anti-neoplastic clinical effects of heparins particularly related to EV uptake [ 53 ]. The benefits of prophylactic or therapeutic heparin medication for thromboembolism or sepsis-induced disseminated intravascular coagulation, particularly related to corona virus disease (COVID19) are currently discussed and investigated in clinical trials [ 55 ].…”

Section: Discussionmentioning

confidence: 99%

Scalable Enrichment of Immunomodulatory Human Acute Myeloid Leukemia Cell Line-Derived Extracellular Vesicles

Binder

Maeding

Wolf

et al. 2021

Cells

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Acute myeloid leukemia (AML) cells can secrete trophic factors, including extracellular vesicles (EVs), instructing the stromal leukemic niche. Here, we introduce a scalable workflow for purification of immunomodulatory AML-EVs to compare their phenotype and function to the parental AML cells and their secreted soluble factors. AML cell lines HL-60, KG-1, OCI-AML3, and MOLM-14 released EVs with a peak diameter of approximately 80 nm in serum-free particle-reduced medium. We enriched EVs >100x using tangential flow filtration (TFF) and separated AML-derived soluble factors and cells in parallel. EVs were characterized by electron microscopy, immunoblotting, and flow cytometry, confirming the double-membrane morphology, purity and identity. AML-EVs showed significant enrichment of immune response and leukemia-related pathways in tandem mass-tag proteomics and a significant dose-dependent inhibition of T cell proliferation, which was not observed with AML cells or their soluble factors. Furthermore, AML-EVs dose-dependently reduced NK cell lysis of third-party K-562 leukemia targets. This emphasizes the peculiar role of AML-EVs in leukemia immune escape and indicates novel EV-based targets for therapeutic interventions.

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Section: Discussionmentioning

confidence: 99%

Scalable Enrichment of Immunomodulatory Human Acute Myeloid Leukemia Cell Line-Derived Extracellular Vesicles

Binder

Maeding

Wolf

et al. 2021

Cells

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“…Human primary keratinocytes (KCs) and fibroblasts (FBs) were obtained from expanded human skin grafts, cultivated in CntPrime medium (CnT-PR, CELLnTEC, Bern, Switzerland) and α-MEM (Sigma-Aldrich) supplemented with 10% HPL, respectively, as described [ 13 ]. Endothelial colony-forming cells (ECFCs) were obtained from human umbilical cord blood as published [ 35 ] and cultured in EGM basal medium (CC-3156, Lonza Group, Basel, Switzerland) supplemented with the bullet kit (EGM-2 medium, CC4176, Lonza) with 10% HPL instead of fetal bovine serum (FBS) including 2 IU/mL heparin to produce endothelial cells (ECs) [ 36 ]. Fibrosphere formation was induced by adding 5,000 FBs per well in a 96-well ultra-low attachment plate (Nunclon Sphera, ThermoFisher Scientific) and skin-organoids were created by mixing KCs, FBs and ECFCs at a 2:1:1 ratio, respectively, totaling 5000 cells/well for quantification or 50,000 cells/well for immunohistochemistry [ 13 ].…”

Section: Methodsmentioning

confidence: 99%

Synergy of Human Platelet-Derived Extracellular Vesicles with Secretome Proteins Promotes Regenerative Functions

et al. 2022

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Platelet-rich plasma is a promising regenerative therapeutic with controversial efficacy. We and others have previously demonstrated regenerative functions of human platelet lysate (HPL) as an alternative platelet-derived product. Here we separated extracellular vesicles (EVs) from soluble factors of HPL to understand the mode of action during skin-organoid formation and immune modulation as model systems for tissue regeneration. HPL-EVs were isolated by tangential-flow filtration (TFF) and further purified by size-exclusion chromatography (SEC) separating EVs from (lipo)protein-enriched soluble fractions. We characterized samples by tunable resistive pulse sensing, western blot, tandem mass-tag proteomics and super-resolution microscopy. We evaluated EV function during angiogenesis, wound healing, organoid formation and immune modulation. We characterized EV enrichment by TFF and SEC according to MISEV2018 guidelines. Proteomics showed three major clusters of protein composition separating TSEC-EVs from HPL clustering with TFF soluble fractions and TFF-EVs clustering with TSEC soluble fractions, respectively. HPL-derived TFF-EVs promoted skin-organoid formation and inhibited T-cell proliferation more efficiently than TSEC-EVs or TSEC-soluble fractions. Recombining TSEC-EVs with TSEC soluble fractions re-capitulated TFF-EV effects. Zeta potential and super-resolution imaging further evidenced protein corona formation on TFF-EVs. Corona depletion on SEC-EVs could be artificially reconstituted by TSEC late fraction add-back. In contrast to synthetic nanoparticles, which commonly experience reduced function after corona formation, the corona-bearing EVs displayed improved functionality. We conclude that permissive isolation technology, such as TFF, and better understanding of the mechanism of EV corona function are required to realize the complete potential of platelet-based regenerative therapies.

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“…In addition, composite nanoparticles can be prepared by mixing oppositely charged polymer solutions with polypositive/anionic solutions. These composite heparin-containing nanoparticles can be combined with high porosity nanostructures, such as chitosan, for binding, stabilization, and release of GFs ( Laner-Plamberger et al, 2021 ).…”

Section: Heparin In Biomaterials Designmentioning

confidence: 99%

The Auxiliary Role of Heparin in Bone Regeneration and its Application in Bone Substitute Materials

Wang

Xiao

Wang

et al. 2022

Front. Bioeng. Biotechnol.

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Bone regeneration in large segmental defects depends on the action of osteoblasts and the ingrowth of new blood vessels. Therefore, it is important to promote the release of osteogenic/angiogenic growth factors. Since the discovery of heparin, its anticoagulant, anti-inflammatory, and anticancer functions have been extensively studied for over a century. Although the application of heparin is widely used in the orthopedic field, its auxiliary effect on bone regeneration is yet to be unveiled. Specifically, approximately one-third of the transforming growth factor (TGF) superfamily is bound to heparin and heparan sulfate, among which TGF-β1, TGF-β2, and bone morphogenetic protein (BMP) are the most common growth factors used. In addition, heparin can also improve the delivery and retention of BMP-2 in vivo promoting the healing of large bone defects at hyper physiological doses. In blood vessel formation, heparin still plays an integral part of fracture healing by cooperating with the platelet-derived growth factor (PDGF). Importantly, since heparin binds to growth factors and release components in nanomaterials, it can significantly facilitate the controlled release and retention of growth factors [such as fibroblast growth factor (FGF), BMP, and PDGF] in vivo. Consequently, the knowledge of scaffolds or delivery systems composed of heparin and different biomaterials (including organic, inorganic, metal, and natural polymers) is vital for material-guided bone regeneration research. This study systematically reviews the structural properties and auxiliary functions of heparin, with an emphasis on bone regeneration and its application in biomaterials under physiological conditions.

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Heparin and Derivatives for Advanced Cell Therapies

Cited by 10 publications

References 179 publications

Scalable Enrichment of Immunomodulatory Human Acute Myeloid Leukemia Cell Line-Derived Extracellular Vesicles

Scalable Enrichment of Immunomodulatory Human Acute Myeloid Leukemia Cell Line-Derived Extracellular Vesicles

Synergy of Human Platelet-Derived Extracellular Vesicles with Secretome Proteins Promotes Regenerative Functions

The Auxiliary Role of Heparin in Bone Regeneration and its Application in Bone Substitute Materials

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