GMP-conformant on-site manufacturing of a CD133+ stem cell product for cardiovascular regeneration

Skorska, Anna; Müller, Paula; Gaebel, Ralf; Große, Jana; Lemcke, Heiko; Lux, Cornelia A.; Bastian, Manuela; Hausburg, Frauke; Zarniko, Nicole; Bubritzki, Sandra; Ruch, Ulrike; Tiedemann, Gudrun; Dávid, Róbert; Steinhoff, Gustav

doi:10.1186/s13287-016-0467-0

Cited by 19 publications

(12 citation statements)

References 34 publications

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“…In the present study, we have characterized the phenotype of both stem cells types. The flow cytometric strategy relied on our previous work [13,15]. Cultivated MSC are often characterized by the expression of several surface marker including CD73, CD105, and CD44 as well as the absence of others, including CD45 and CD34 [2,13].…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, we have examined whether CD133 + and CD271 + cells form separate populations within BM or if the surface proteins are also expressed simultaneously by an overlap population, which is an important aspect in order to evaluate the feasibility of a combined therapeutic approach. Taking into account that the majority of CD133 + cells express CD34 + , MACS-purified CD133 + cells were co-stained with CD34 antibody [14,15]. Moreover, the CD271 + population was co-stained for CD45 for further distinguishing of the MSC subpopulation lacking the CD45 surface marker [16].…”

Section: Flow Cytometric Characterization Of Stem Cell Populations Qmentioning

confidence: 99%

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Angiogenic Potential of Bone Marrow Derived CD133+ and CD271+ Intramyocardial Stem Cell Trans- Plantation Post MI

Sasse

Skorska

Lux

et al. 2019

Cells

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Background: Bone marrow (BM)-derived stem cells with their various functions and characteristics have become a well-recognized source for the cell-based therapies. However, knowledge on their therapeutic potential and the shortage for a cross-link between distinct BM-derived stem cells, primed after the onset of myocardial infarction (MI), seems to be still rudimentary. Therefore, the post-examination of the therapeutic characteristics of such primed hematopoietic CD133+ and mesenchymal CD271+ stem cells was the object of the present study. Methods and Results: The effects of respective CD133+ and CD271+ mononuclear cells alone as well as in the co-culture model have been explored with focus on their angiogenic potential. The phenotypic analysis revealed a small percentage of isolated cells expressing both surface markers. Moreover, target stem cells isolated with our standardized immunomagnetic isolation procedure did not show any negative alterations following BM storage in regard to cell numbers and/or quality. In vitro network formation relied predominantly on CD271+ stem cells when compared with single CD133+ culture. Interestingly, CD133+ cells contributed in the tube formation, only if they were cultivated in combination with CD271+ cells. Additional to the in vitro examination, therapeutic effects of the primed stem cells were investigated 48 h post MI in a murine model. Hence, we have found a lower expression of transforming growth factor βeta 3 (TGFβ3) as well as an increase of the proangiogenic factors after CD133+ cell treatment in contrast to CD271+ cell treatment. On the other hand, the CD271+ cell therapy led to a lower expression of the inflammatory cytokines. Conclusion: The interactions between CD271+ and CD133+ subpopulations the extent to which the combination may enhance cardiac regeneration has still not been investigated so far. We expect that the multiple characteristics and various regenerative effects of CD271+ cells alone as well as in combination with CD133+ will result in an improved therapeutic impact on ischemic heart disease.

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Section: Discussionmentioning

confidence: 99%

Section: Flow Cytometric Characterization Of Stem Cell Populations Qmentioning

confidence: 99%

Angiogenic Potential of Bone Marrow Derived CD133+ and CD271+ Intramyocardial Stem Cell Trans- Plantation Post MI

Sasse

Skorska

Lux

et al. 2019

Cells

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“…Some have been defined as pro-arrhythmogenic cells, i.e., skeletal myoblasts, while others have shown antiarrhythmic effects, such as embryonic cardiomyocytes and mesenchymal stem cells (MSC) [2][3][4][5]. Our research group has focused on the role of human bone marrow (BM) derived hematopoiesis such as MSC in regenerative medicine [6][7][8][9]. In previous studies we have shown that it is safe to use CD117 + stem cells for myocardial regeneration, as their electrophysiological properties…”

Section: Introductionmentioning

confidence: 99%

CD271+ Human Mesenchymal Stem Cells Show Antiarrhythmic Effects in a Novel Murine Infarction Model

Sadraddin

Gaebel

Skorska

et al. 2019

Cells

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Background: Ventricular arrhythmias (VA) are a common cause of sudden death after myocardial infarction (MI). Therefore, developing new therapeutic methods for the prevention and treatment of VA is of prime importance. Methods: Human bone marrow derived CD271 + mesenchymal stem cells (MSC) were tested for their antiarrhythmic effect. This was done through the development of a novel mouse model using an immunocompromised Rag2 −/− γc −/− mouse strain subjected to myocardial "infarction-reinfarction". The mice underwent a first ischemia-reperfusion through the left anterior descending (LAD) artery closure for 45 min with a subsequent second permanent LAD ligation after seven days from the first infarct. Results: This mouse model induced various types of VA detected with continuous electrocardiogram (ECG) monitoring via implanted telemetry device. The immediate intramyocardial delivery of CD271 + MSC after the first MI significantly reduced VA induced after the second MI. Conclusions: In addition to the clinical relevance, more closely reflecting patients who suffer from severe ischemic heart disease and related arrhythmias, our new mouse model bearing reinfarction warrants the time required for stem cell engraftment and for the first time enables us to analyze and verify significant antiarrhythmic effects of human CD271 + stem cells in vivo. make them very unlikely to produce hazardous action potentials or contribute to arrhythmias [10,11]. However, the availability of a realistic small animal model reflecting the situation in patients is of utmost importance. For testing the rhythmological behavior of human cells, Rag2 −/− γc −/− has been selected. These mice (strain C, 129S4-Rag2 tm1.1Flv Il2rg tm1.1Flv /J) have been derived from a V17 embryonic stem cell line (BALB/c × 129 heterozygote) targeted for the Rag2 and Il2rg genes and lack B, T, and natural killer (NK) cells [12]. They have a deletion of the common cytokine receptor γ chain (γc) gene, and thus reduced numbers of peripheral T and B lymphocytes, and absent natural killer cell (NK) activity. A genetic cross with a recombinase activating gene 2 (RAG2) deficient strain produced mice doubly homozygous for the γc and RAG2 null alleles (Rag2 −/− γc −/− ) with a stable phenotype characterized by the absence of all T lymphocyte, B lymphocyte, and NK cell function [13]. This phenotype makes the completely a lymphoid strain useful for studies on human tissue xenotransplantation [14].As previously shown, stem cell antiarrhythmic effects appear after completion of an approximately one-week engraftment period [4]. The major objective of the present study was to develop a mouse model that enables us to reproduce VA several days after an initial MI. In vivo electrophysiological mouse models are well established: Previously, Berul et al. established an open chest epicardial study of conduction properties with the ability to induce second and third degree heart blocks, but no induction of tachyarrhythmias after programmed stimulation [15]. Hagendorff et al. used a rapid t...

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“…With numbers of 10 9 MSC, effects were reported in treatment of some autoimmune and infectious diseases [68,69]. A lot of clinical trials have been performed or are ongoing on treating cardiovascular diseases (myocardial infarction, cardiomyopathy, critical limb ischemia, stroke [72][73][74]). Single or repeated doses of 10 7 to more than 10 9 are injected systemically or topically.…”

Section: State-of-the-art With Ex Vivo Expanded Mscs Utilized In Clinmentioning

confidence: 99%

Landscape of Manufacturing Process of ATMP Cell Therapy Products for Unmet Clinical Needs

Pörtner¹,

Parida²,

Schaffer³

et al. 2018

Stem Cells in Clinical Practice and Tissue Engineering

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Immune cell therapies have been studied in numerous clinical trials using Advanced Therapy Medicinal Products (ATMP) against a number of diseases having no or inadequate alternative therapies available, for example, various cancer types, cerebral stroke, cardiac infarction, severe autoimmune disorders, or chronic infections. Despite the enormous number of positive observation in ex vivo or animal studies, convincing results in clinical studies remain scanty. The chapter presents a survey and reveals that the manufacturing of immune cells especially for clinical trials is until today primarily performed using archaic, scarcely controlled, and incomparable processes and methods. A deeper characterization of ex vivo expanded immune cells is urgently needed not only on the level of a few receptors and ligands on the cell surface but also with respect to the ever-contained subtypes in an expanded immune cell population, the pattern of secreted effector molecules, and their amounts over time and influences from in vivo components on them.

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GMP-conformant on-site manufacturing of a CD133+ stem cell product for cardiovascular regeneration

Cited by 19 publications

References 34 publications

Angiogenic Potential of Bone Marrow Derived CD133+ and CD271+ Intramyocardial Stem Cell Trans- Plantation Post MI

Angiogenic Potential of Bone Marrow Derived CD133+ and CD271+ Intramyocardial Stem Cell Trans- Plantation Post MI

CD271+ Human Mesenchymal Stem Cells Show Antiarrhythmic Effects in a Novel Murine Infarction Model

Landscape of Manufacturing Process of ATMP Cell Therapy Products for Unmet Clinical Needs

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