Cellular therapy to treat heart failure is an ongoing focus of intense research, but progress toward structural and functional recovery remains modest. Engineered augmentation of established cellular effectors overcomes impediments to enhance reparative activity. Such 'next generation' implementation includes delivery of combinatorial cell populations exerting synergistic effects. Concurrent isolation and expansion of three distinct cardiac-derived interstitial cell types from human heart tissue, previously reported by our group, prompted design of a 3D structure that maximizes cellular interaction, allows for defined cell ratios, controls size, enables injectability, and minimizes cell loss. Herein, mesenchymal stem cells (MSCs), endothelial progenitor cells (EPCs) and c-Kit + cardiac interstitial cells (cCICs) when cultured together spontaneously form scaffold-free 3D microenvironments termed Cardi-oClusters. scRNA-Seq profiling reveals CardioCluster expression of stem cell-relevant factors, adhesion/extracellular-matrix molecules, and cytokines, while maintaining a more native transcriptome similar to endogenous cardiac cells. CardioCluster intramyocardial delivery improves cell retention and capillary density with preservation of cardiomyocyte size and long-term cardiac function in a murine infarction model followed 20 weeks. CardioCluster utilization in this preclinical setting establish fundamental insights, laying the framework for optimization in cell-based therapeutics intended to mitigate cardiomyopathic damage.
1Background: Cellular therapy to treat heart failure is an ongoing focus of intense 2 research and development, but progress has been frustratingly slow due to limitations of 3 current approaches. Engineered augmentation of established cellular effectors 4 overcomes impediments, enhancing reparative activity with improved outcomes relative 5 to conventional techniques. Such 'next generation' implementation includes delivery of 6 combinatorial cell populations exerting synergistic effects. Concurrent isolation and 7 expansion of three distinct cardiac-derived interstitial cell types from human heart tissue, 8 as previously reported by our group, prompted design of a three-dimensional (3D) 9 structure that maximizes cellular interaction, allows for defined cell ratios, controls size, 10 enables injectability, and minimizes cell losses upon delivery. 11 Methods: Three distinct populations of human cardiac interstitial cells including 12 mesenchymal stem cells (MSCs), endothelial progenitor cells (EPCs), and c-Kit + cardiac 13 interstitial cells (cCICs) when cultured together spontaneously form scaffold-free 3D 14 microenvironments termed CardioClusters. Biological consequences of CardioCluster 15 formation were assessed by multiple assays including single cells RNA-Seq 16 transcriptional profiling. Protective effects of CardioClusters in vitro were measured 17 using cell culture models for oxidative stress and myocardial ischemia in combination 18 with freshly isolated neonatal rat ventricular myocytes. Long-term impact of adoptively 19 transferred CardioClusters upon myocardial structure and function in a xenogenic model 20 of acute infarction using NOD scid mice was assessed over a longitudinal time course of 21 20-weeks. 22 Results: CardioCluster design enables control over composite cell types, cell ratios, 1 size, and preservation of structural integrity during delivery. Profound changes for 2 biological properties of CardioClusters relative to constituent parental cell populations 3 include enhanced expression of stem cell-relevant factors, adhesion/extracellular-matrix 4 molecules, and cytokines. The CardioCluster 3D microenvironment maximizes cellular 5 interaction while maintaining a more native transcriptome similar to endogenous cardiac 6 cells. CardioCluster delivery improves cell retention following intramyocardial injection 7 with preservation of long-term cardiac function relative to monolayer-cultured cells when 8 tested in an experimental murine infarction model followed for up to 20 weeks post-9 challenge. CardioCluster-treated hearts show increases in capillary density, 10 preservation of cardiomyocyte size, and reduced scar size indicative of blunting 11 pathologic infarction injury. 12 Conclusions: CardioClusters are a novel 'next generation' development and delivery 13 approach for cellular therapeutics that potentiate beneficial activity and enhance 14 protective effects of human cardiac interstitial cell mixed populations. CardioClusters 15 utilization in this preclinical setting establishes ...
Cardiac interstitial cells (CICs) perform essential roles in myocardial biology through preservation of homeostasis as well as response to injury or stress. Studies of murine CIC biology reveal remarkable plasticity in terms of transcriptional reprogramming and ploidy state with important implications for function. Despite over a decade of characterization and in vivo utilization of adult c-Kit + CIC (cCIC), adaptability and functional responses upon delivery to adult mammalian hearts remain poorly understood. Limitations of characterizing cCIC biology following in vitro expansion and adoptive transfer into the adult heart were circumvented by delivery of the donated cells into early cardiogenic environments of embryonic, fetal, and early postnatal developing hearts. These three developmental stages were permissive for retention and persistence, enabling phenotypic evaluation of in vitro expanded cCICs after delivery as well as tissue response following introduction to the host environment. Embryonic blastocyst environment prompted cCIC integration into trophectoderm as well as persistence in amniochorionic membrane. Delivery to fetal myocardium yielded cCIC perivascular localization with fibroblast-like phenotype, similar to cCICs introduced to postnatal P3 heart with persistent cell cycle activity for up to 4 weeks. Fibroblast-like phenotype of exogenously transferred cCICs in fetal and postnatal cardiogenic environments is consistent with inability to contribute directly toward cardiogenesis and lack of functional integration with host myocardium. In contrast, cCICs incorporation into extraembryonic membranes is consistent with fate of polyploid cells in blastocysts. These findings provide insight into cCIC biology, their inherent predisposition toward fibroblast fates in cardiogenic environments, and remarkable participation in extraembryonic tissue formation.
2 Cardiac interstitial cells (CIC) perform essential roles in myocardial biology through 3 preservation of homeostasis as well as response to injury or stress. Studies of murine 4 CIC biology reveal remarkable plasticity in terms of transcriptional reprogramming and 5 ploidy state with important implications for function. Despite over a decade of 6 characterization and in vivo utilization of adult c-Kit + CIC (cCIC), adaptability and 7 functional responses upon delivery to adult mammalian hearts remain poorly understood. 8 Limitations of characterizing cCIC biology following in vitro expansion and adoptive 9 transfer into the adult heart were circumvented by delivery of the donated cells into early 10 cardiogenic environments of embryonic, fetal, and early postnatal developing hearts.11
Introduction: Stem cell therapy represents great promises to myocardium regeneration. Multipotent c-Kit pos cardiac progenitor cells (CPCs) are able to differentiate into endothelial cells, smooth muscle cells, and cardiomyocytes. However, fundamental knowledge of CPC biology remains incomplete. Studies in rodent myocardial infarction model revealed that CPCs have poor long-term survival and engraftment after adoptive transfer, perhaps due to the severely damaged host environment. Therefore, it is critical to understand how CPCs interface with the recipient environment following transfer in order to enhance their true regenerative potentials. Hypothesis: Adoptively transferred stem cells are thought to survive and engraft best in an environment closely resembling their original habitat. Thus, we hypothesized that the embryonic environment provides the optimal spatiotemporal conditions to promote CPCs engraftment and commitment to cardiac fate. Methods: CPCs isolated from adult mouse hearts were expanded, fluorescence-tagged, and injected into blastocysts at E3.75 and in utero at E15.5. Embryos were analyzed following cardiogenesis by immunofluorescence for presence of CPC-derived tissues. Additionally, CPCs were injected intramyocardially at various stages from P0 to P7, to follow long-term adoptive transfer and assess CPCs lineage commitment. Results and Conclusions: At 48 hours post injection, donor CPCs were found anchoring in blastocoel and trophoblasts at E5.5, and were detected within the host myocardium at E17.5 predominantly at perivascular regions (n=4). Interestingly, CPCs also integrated into aminochorionic sac, indicating a novel non-cardiogenic fate of CPCs (n=5). CPCs injected at P3 stably engrafted into left ventricular myocardium by 14 days post injection (n=4), sharing gap junction proteins (ZO-1, Connexin-43) with neighboring cells. In conclusion, this study provides vivid evidence for the first time of CPC engraftment and survival in vivo under homeostasis during cardiogenesis. Future studies will assess the permissive environmental conditions, which may optimize their use in therapeutic applications, and the cardiogenic potential of CPCs in order to provide fundamental insights on CPCs biology.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.