Donor Cells Research Articles

Direct Reprogramming of Somatic Skin Cells from a Patient with Huntington’s Disease into Striatal Neurons to Create Models of Pathology

A new in vitro model of Huntington’s disease (HD) was developed via a direct reprogramming of dermal fibroblasts from HD patients into striatal neurons. A reprogramming into induced pluripotent stem (iPS) cells is obviated in the case of direct reprogramming, which thus yields neurons that preserve the epigenetic information inherent in cells of a particular donor and, consequently, the age-associated disease phenotype. A main histopathological feature of HD was reproduced in the new model; i.e., aggregates of mutant huntingtin accumulated in striatal neurons derived from a patient’s fibroblasts. Experiments with cultured neurons obtained via direct reprogramming make it possible to individually assess the progression of neuropathology and to implement a personalized approach to choosing the treatment strategy and drugs for therapy. The in vitro model of HD can be used in preclinical drug studies.

Donor-Derived Engineered Microvessels for Cardiovascular Risk Stratification of Patients with Kidney Failure.

Cardiovascular disease is the cause of death in ≈50% of hemodialysis patients. Accumulation of uremic solutes in systemic circulation is thought to be a key driver of the endothelial dysfunction that underlies elevated cardiovascular events. A challenge in understanding the mechanisms relating chronic kidney disease to cardiovascular disease is the lack of in vitro models that allow screening of the effects of the uremic environment on the endothelium. Here, a method is described for microfabrication of human blood vessels from donor cells and perfused with donor serum. The resulting donor-derived microvessels are used to quantify vascular permeability, a hallmark of endothelial dysfunction, in response to serum spiked with pathophysiological levels of indoxyl sulfate, and in response to serum from patients with chronic kidney disease and from uremic pigs. The uremic environment has pronounced effects on microvascular integrity as demonstrated by irregular cell-cell junctions and increased permeability in comparison to cell culture media and healthy serum. Moreover, the engineered microvessels demonstrate an increase in sensitivity compared to traditional 2D assays. Thus, the devices and the methods presented here have the potential to be utilized to risk stratify and to direct personalized treatments for patients with chronic kidney disease.

Recruiting refugees and migrants as potential hematopoietic stem cell donors to serve patients of comparable ethnicities with rare human leucocyte antigen patterns - The BluStar.NRW project in North Western Germany

Currently, approximately 19 million people with a migration background live in Germany. The majority of those descend from regions where the population has a genetically different distribution of HLA antigens when compared to the HLA frequencies usually found in North Western Europe. In case of severe haematological disorders of these individuals, allogeneic stem cell transplantation may be the treatment of choice. However, finding appropriate histocompatible hematopoietic stem cell donors continues to be a major challenge. If no matching sibling donors are available, there are only few suitable donors with a similar genetic background available in international blood stem cell donor registries.The “BluStar.NRW” project aimed to recruit new blood and hematopoietic stem cell donors with a migration background and to noticeably increase the number of suitable donors for patients within this group. Since December 2017, a total number of 9100 blood and stem cell donors with a migration background were recruited and typed for this project. HLA typing for HLA-A, -B, -C, -DRB1, -DQB1, and -DPB1 was performed by Next Generation Sequencing. We assessed the proportion of rare alleles according to HLA frequency tables, as defined by a frequency of <1:1000. The rare HLA allele frequencies according to HLA frequency tables of the BluStar.NRW cohort were compared with a matched control donor cohort: Rare HLA-A, -B, -C, -DRB1 and -DQB1 alleles occurred three times more frequent than in the control group, but rare HLA-DPB1 alleles occurred more frequently in the control cohort. This difference was highly significant for all HLA alleles (p < 0.0001 for HLA-A, -B, -C, -DRB1, -DPB1; p = 0.0002 for HLA-DQB1). In addition, the distribution of rare alleles differed between the two groups. To date, 29 work-ups were initiated, 12 PBSC, one BM and three DLI were collected so far out of the BluStar.NRW cohort. The apheresis probability is twofold higher (0.18% vs. 0.07%) compared to the control group which clearly shows a serious medical need. However, 13 work-ups were cancelled in the BluStar.NRW donor cohort which represents an almost twice as higher cancellation rate (45% vs. 25%).This single registry analysis with a large sample cohort clearly indicates that hematopoietic stem cell donors with a migration background represent an adequate donor pool to serve patients of comparable ethnicity.

Biogenesis and delivery of extracellular vesicles: harnessing the power of EVs for diagnostics and therapeutics.

Extracellular vesicles (EVs) are membrane-enclosed particles secreted by a variety of cell types. These vesicles encapsulate a diverse range of molecules, including proteins, nucleic acids, lipids, metabolites, and even organelles derived from their parental cells. While EVs have emerged as crucial mediators of intercellular communication, they also hold immense potential as both biomarkers and therapeutic agents for numerous diseases. A thorough understanding of EV biogenesis is crucial for the development of EV-based diagnostic developments since the composition of EVs can reflect the health and disease status of the donor cell. Moreover, when EVs are taken up by target cells, they can exert profound effects on gene expression, signaling pathways, and cellular behavior, which makes these biomolecules enticing targets for therapeutic interventions. Yet, despite decades of research, the intricate processes underlying EV biogenesis by donor cells and subsequent uptake by recipient cells remain poorly understood. In this review, we aim to summarize current insights and advancements in the biogenesis and uptake mechanisms of EVs. By shedding light on the fundamental mechanisms governing EV biogenesis and delivery, this review underscores the potential of basic mechanistic research to pave the way for developing novel diagnostic strategies and therapeutic applications.

Impact of COVID-19 pandemic on cord blood banking and transplantation.

Umbilical cord blood is a rich source of hematopoietic stem cells that has been used for transplantation for over 30 years, especially when there is no compatible hematopoietic stem cell donor available. Its use has decreased more recently, since the development of methods to improve haploidentical transplants has allowed the use of mobilized peripheral blood as a source of hematopoietic stem cells. Public cord blood banks collect, process and store cord blood samples from voluntary donations. In addition, many public banks are involved in research to enhance hematopoietic stem cell therapies and develop new treatments for haematological and genetic diseases. The COVID-19 pandemic, which emerged in 2019, has had a profound and wide-ranging impact on human health and treatment. The area of hematopoietic stem cell transplantation was deeply affected by reductions in bone marrow, peripheral blood and cord blood donations; logistical challenges; exposure of healthcare workers and other challenges. The present study reviews the impact of the COVID-19 pandemic on cord blood banking and transportation around the world with a special focus on Brazil.

SHEDDING LIGHT ON CHONDROGENESIS: THE IMPACT OF LOW-LEVEL LASER ON THE CHONDROGENIC DIFFERENTIATION

Photobiomodulation (PBM), the use of light for regenerative purposes, has a long history with first documentations several thousand years ago in ancient Egypt and a Nobel Price on this topic at the beginning of last century (by Niels Finsen). Nowadays, it is in clinical use for indications such as wound healing, pain relief and anti-inflammatory treatment. Given the rising numbers of in vitro studies, there is increasing evidence for the underlying mechanisms such as wavelength dependent reactive oxygen production and adenosine triphosphate generation. In cartilage regeneration, the use of PBM is controversially discussed with divergent results in clinics and insufficient in vitro studies. As non-invasive therapy, PMB is, though, of particular importance, since a general regenerative stimulus would be of great benefit in the otherwise only surgically accessible tissues. We therefore investigated the influence of different wavelengths - blue (475 nm), green (516 nm) or red (635 nm) of a low-level laser (LLL) - on the chondrogenic differentiation of chondrocytes and adipose derived stromal cells of different human donors and applied the light in different settings (2D, 3D) with cells in a proliferative or differentiating stage. All assessed parameters (spheroid growth, histology, matrix quantification and gene expression) revealed an influence of LLL on chondrogenesis in a donor-, wavelength- and culture-model-dependent manner. Especially encouraging was the finding, that cells with poor chondrogenic potential could be improved by one single 2D treatment. Amongst the three wave lengths, red light was the most promising one with the most positive impact. Although in vivo data are still missing, these in vitro results provide evidence for a proper biofunctional effect of LLL.

Alterations in clinical parameters in cicatrising conjunctivitis secondary to GvHD

Cicatricial conjunctivitis is part of a well‐known complication of graft versus host disease (GVHD) resulting from the tissue fibrosis that affects different body organs. It results from inflammatory reactions determined by recognition of recipient antigens as foreign by donor cells. The main target tissues in ocular GVHD are the conjunctiva and the lacrimal gland, and conjunctival involvement is usually associated with more severe systemic disease. Clinical manifestations of cicatricial conjunctivitis includes conjunctival fibrosis, conjunctival shortening, conjunctival scarring and keratinization, and symblepharon and/or ankyloblepharon. Such scarring may even lead to cicatricial lagophthalmos, ectropion, entropion, and stenosis or closure of the lacrimal puncta. In fact, the degree of palpebral conjunctival changes has been used as a criterion to classify conjunctival involvement in chronic GVHD.Conjunctival epithelial cells and myofibroblasts may have donor origin, though with a very small proportion. These donor‐derived fibroblasts may play a role in the pathogenesis of conjunctival fibrosis. In addition, it has been shown that the ocular surface of patients with chronic GVHD may show epithelial‐mesenchymal transition, in which the epithelium may gain the mesenchymal phenotype and the ability to migrate into the subepithelial stroma, resulting in conjunctival fibrosis.Our group showed that a significant proportion of patients with ocular GHVD had subtarsal fibrosis, that was non evident in non‐GHVD dry eye disease (DED) patients, even with the same degree of clinical severity. Extent of involvement varied from one eye to another; on average, more than a quarter of the visible tarsal area was affected by the fibrosis. Such fibrosis was significantly associated with ocular surface epitheliopathy and conjunctival staining. In contrast, densities of epithelial and stromal immune cells in the conjunctiva were comparable between GVHD and non GVHD DED patients after adjustment for severity of DED. We also showed that spontaneous extrusion of punctal plugs occurs in a majority of ocular GVHD‐associated DED patients within 90 days of plug placement, and this lower retention rate of punctal plugs has been attributed to punctal subepithelial fibrosis.In all patients with ocular GVHD, the eye care provider should evaluate the presence of conjunctival fibrosis and associated ocular surface complications. If present, the therapeutic strategy should attempt to address this scarring process.

Dependence of the reproduction coefficient on donor cells in in vitro improvement of potato cultivars

In the experiment, along with growing potato varieties from the meristem cells of the tuber growth, we germinate the botanical seeds of the varieties, grow them from the growth, grow the primary donor cells, obtain healthy callus tissue, grow morphogenetically identical cells, the yield of healthy plant sprouts from the callus tissue is 20% compared to the usual experimental options. 25-30% of callus tissue sprouts, which did not meet the requirements and changed in morphogenesis, remained underdeveloped when callus tissue was grown by taking cells from a normal tumor. It was found that the morphogenesis of the callus sprouts, which were germinated from the seed, obtained from the growths and formed, the weight of the remaining sprouts was 9-10%, and the reproduction coefficient of the sprouts was more than 20%.

Small extracellular vesicles: Roles and clinical application in prostate cancer.

Prostate cancer is a significant health problem in the United States. It is remarkably heterogenous, ranging from slow growing disease amenable to active surveillance to highly aggressive forms requiring active treatments. Therefore, being able to precisely determine the nature of disease and appropriately match patients to available and/or novel therapeutics is crucial to improve patients' overall outcome and quality of life. Recently small extracellular vesicles (sEVs), a subset of nanoscale membranous vesicles secreted by various cells, have emerged as important analytes for liquid biopsy and promising vehicles for drug delivery. sEVs contain various biomolecules such as genetic material, proteins, and lipids that recapitulate the characteristics and state of their donor cells. The application of existing and newly developed technologies has resulted in an increased depth of knowledge about biophysical structures, biogenesis, and functions of sEVs. In prostate cancer patients, tumor-derived sEVs can be isolated from biofluids, commonly urine and blood. They mediate intercellular signaling within the tumor microenvironment and distal organ-specific sites, supporting cancer initiation, progression, and metastasis. A mounting body of evidence suggests that sEV components can be potent biomarkers for prostate cancer diagnosis, prognosis, and prediction of disease progression and treatment response. Due to enhanced circulation stability and bio-barrier permeability, sEVs can be also used as effective drug delivery carriers to improve the efficacy and specificity of anti-tumor therapies. This review discusses recent studies on sEVs in prostate cancer and is focused on their role as biomarkers and drug delivery vehicles in the clinical management of prostate cancer.

A case for the study of native extracellular vesicles.

Three main areas of research revolve around extracellular vesicles (EVs): their use as early detection diagnostics for cancer prevention, engineering of EVs or other enveloped viral-like particles for therapeutic purposes and to understand how EVs impact biological processes. When investigating the biology of EVs, it is important to consider strategies able to track and alter EVs directly in vivo, as they are released by donor cells. This can be achieved by suitable engineering of EV donor cells, either before implantation or directly in vivo. Here, we make a case for the study of native EVs, that is, EVs released by cells living within a tissue. Novel genetic approaches to detect intercellular communications mediated by native EVs and profile recipient cells are discussed. The use of Rab35 dominant negative mutant is proposed for functional in vivo studies on the roles of native EVs. Ultimately, investigations on native EVs will tremendously advance our understanding of EV biology and open novel opportunities for therapy and prevention.

Human Leukocyte Antigen Typing in Organ Transplantation: Methods, Clinical Relevance, and Practical Applications

ABSTRACT Transplantation of tissues and organs is one of the greatest achievements of this century. Antigens which vary between members of same species are known as allo-antigens. Difference in the allo-antigens between the donor and the recipient represents the antigenic stimulus, which can cause rejections. Adaptive immunity identifies self from non-self. The main objective of the immune response is to identify the cell surface molecules (major histocompatibility complex /MHC) expressed on the donor cells. It is imperative that human leukocyte antigens (HLA) antigens are identified to gauge the mismatches. Presence of pre formed HLA antibodies or formation of de-novo HLA antibodies against these mismatched antigens can lead to antibody mediated rejections and decreased allograft survival. Identification and monitoring of these antibodies pre transplant and post-transplant by performing a virtual cross-match with mismatched donor antigens help in planning and adjusting immunosuppression. A precise and adequate HLA typing of the donor and recipient is required for virtual cross-match. HLA typing technologies have advanced from serological typing to molecular technologies, which can now help identify the donor tissue to allelic level. Methods of HLA typing and their applications with cases have been described in this article.

Modeling Extracellular Vesicles-Mediated Interactions of Cells in the Tumor Microenvironment.

Interactions of cells via extracellular vesicles (EVs) manipulate various actions, including cancer initiation and progression, inflammation, anti-tumor signaling and cell migration, proliferation and apoptosis in the tumor microenvironment. EVs as the external stimulus can activate or inhibit some receptor pathways in a way that amplify or attenuate a kind of particle release at target cells. This can also be carried out in a biological feedback-loop where the transmitter is affected by the induced release initiated by the target cell due to the EVs received from the donor cell, to create a bilateral process. In this paper, at first we derive the frequency response of internalization function in the framework of a unilateral communication link. This solution is adapted to a closed-loop system to find the frequency response of a bilateral system. The overall releases of the cells, given by the combination of the natural release and the induced release, are reported at the end of this paper and the results are compared in terms of distance between the cells and reaction rates of EVs at the cell membranes.

Hematopoietic stem cell transplantation: an Italian monocentric experience on the health assessment and eligibility of adult-related donors.

Indications for HSCT are increasing worldwide, paralleled by a growing demand for donors of therapeutic cells. Herein, we report our real-world experience of adult HPC donor assessment during a 5-year study period (2018-2023): we have retrospectively revised data of 455 potential related stem cell donors, consecutively evaluated at our center. Donor medical history was assessed by a questionnaire and an interview with a trained physician experienced in donation procedures to evaluate donor fitness and medical history. Pre-existing health disorders were fully investigated. Behavioral risk factors for communicable infectious diseases were also routinely explored. Overall, 351 donors were finally assessed as eligible for HPC donation, and 233 underwent stem cell collection, 158 through apheresis from mobilized peripheral blood, and 75 through bone marrow harvest. Among them, 27 donors were selected despite the presence of pre-existing health conditions, which would be potential exclusion criteria for unrelated donors: 16 suffered from well-controlled cardiovascular diseases (CVD) and 11 from allergic diathesis. Most of the selected donors with pre-existing disorders were candidates for apheresis HPC collection (21, 77.8%), while only six (22.2%) underwent BM harvest. We then analyzed the data relative to the corresponding 233 allogeneic HSCT to explore if the presence of pre-existing diseases in the donors could show any association with transplant characteristics. Transplants from CVD and allergy donors showed no significant disparities in comparison with those from healthy donors. A significant difference emerged regarding the disease severity, with a higher proportion of patients with high/very high disease risk index (DRI) among those receiving grafts from CVD donors (68.7% in transplants from CVD donors versus 36.0% in transplants from healthy donors, p=0.005). Multivariate analysis confirmed that high/very high DRI patients had an increased probability of receiving donations from CVD donors (OR, 4.89; 95%CI, 1.15-20.86; p=0.031). Among donors with well-controlled pre-existing conditions, no adverse events were recorded during stem cell collection or at follow-up. Our results suggest that in patients at high risk for relapse requiring a prompt allogeneic transplant, a familiar donor might be accepted for HPC apheresis donation on less strict criteria than unrelated donors, without risk for both donor and patient.

Antifibrotic activity of carbon quantum dots in a human in vitro model of non-alcoholic steatohepatitis using hepatic stellate cells.

Around 33% of the global population suffers from non-alcoholic fatty liver disease (NAFLD). From these patients, 30% of them progress into non-alcoholic steatohepatitis (NASH), the critical point where lack of treatment leads to cirrhosis and hepatic failure. Moreover, to date, there are no approved therapeutic options available for NASH. It is known that hepatic stellate cell (HSC) activation contributes the most to hepatic disfunction, leading to reactive oxygen species (ROS) accumulation and chronic inflammation, and that the use of nanomaterials to deliver antioxidants may have potential to reduce the activity of activated HSCs. Therefore, we implemented a human in vitro co-culture model in which we take into consideration two factors related to NASH and fibrosis: human hepatic stellate cells from a NASH diagnosed donor (HHSC-N) and peripheral blood mononuclear cells (PBMCs), particularly lymphocytes. The co-cultures were treated with: (1) carbon quantum dots (CD) or (2) lactoferrin conjugated CD (CD-LF) for 24 h or 72 h. CD and CD-LF treatments significantly downregulated profibrotic genes' expression levels of ACTA2, COL1A1, and TIMP1 in co-cultured HHSC-N at 72 h. Also, we assayed the inflammatory response by quantifying the concentrations of cytokines IL-22, IL-10, IFN-γ and IL-4 present in the co-culture's conditioned media whose concentrations may suggest a resolution-associated response in progress. Our findings may serve as a starting point for the development of a NASH treatment using bio-nanotechnology.

Pretransfusion Testing and Daratumumab: Assessing Interference Risks and Developing Solutions

ABSTRACT Daratumumab is a monoclonal antibody; anti-CD38 is used for the treatment of multiple myeloma and other hematological disorders. Daratumumab, binds to CD 38 on red blood cells (RBCs), interfering with pretransfusion testing. This interference can lead to cross match incompatibility and panreactive indirect Antiglobulin tests (IAT).Panreactive IAT can mask presence of underlying clinically significant alloantibodies, especially in patient with history of multiple blood transfusions. This can lead to delays in transfusion, increased risk of transfusion reaction and potential risk for transfusion recipients. Daratumumab induced potential serological interference needs to be resolved by using appropriate methodologies like dithiotheritol (DDT) and enzymatic treatment of reagents or donors cells to overcome these incompatibilities. Effective communication between clinical practitioners and blood centre before starting daratumumab therapy can prevent unnecessary workload and delay in transfusion. Here, we describe the case of interference in prretransfuison testing caused by Daratumumab in the red cell compatibility testing of a patient with severe aplastic anemia and strongly positive DSA levels.

Enriching new transplantable RGC-like cells from retinal organoids for RGC replacement therapy

Optic neuropathies, such as glaucoma, are due to progressive retinal ganglion cells (RGCs) degeneration, result in irreversible vision loss. The promising RGCs replacement therapy for restoring vision are impeded by insufficient RGC-like cells sources. The present work was enriched one new type RGC-like cells using two surface markers CD184 and CD171 from human induced pluripotent stem cells (hiPSCs) by FACS sorting firstly. These new kind cells have well proliferation ability and possessed passage tolerance in vitro 2D or 3D spheroids culture, which kept expressing Pax6, Brn3b and βIII-Tubulin and so on. The transplanted CD184+CD171+ RGC-like cells could survive and integrate into the normal and optic nerve crush (ONC) mice retina, especially they were more inclined to across the optic nerve head and extend to the damaged optic nerve. These data support the feasible application for cell replacement therapy in RGC degenerative diseases, as well as help to develop new commercial cells sorting reagents and establish good manufacturing practice (GMP) grade RGC-like donor cells for further clinical application.

HLA-B*51 Frequency in Transplant Patients and Donors.

HLA molecules play a crucial role in transplantation. The best treatment modality in patients with end-stage renal disease is renal transplant. HLA mismatches between patients and donors can prolong time for renal transplant therapy, reduce graft survival, and increase mortality. HLA region is the most polymorphic genetic region and is essential for antigen presentation. The main target of the recipient's immune system is HLA molecules on the surface of donor cells. HLA-B*51 is associated with Behcet disease, a rare multisystemic disease characterized by autoimmunity and inflammatory processes. In transplant recipients, inflammation and vasculitis are immunologic mechanisms that are responsible for damage of graft tissue. In this retrospective study, we aimed to investigate the frequency of HLA-B*51 in patients diagnosed with end-stage renal disease and in controls and to investigate correlations with rejection episodes. Patients who applied to Baskent University Adana Dr. Turgut Noyan Research and Medical Center (between 2010 and 2022) with end-stage renal disease (n = 1732) and a control group (n = 5277) received HLA typing for class I (HLA-A, HLA-B). Sequence-specific primers or sequencespecific oligonucleotides were used. Among patients diagnosed with end-stage renal disease, 321 had kidney transplant. Frequency of HLA-B*51 was 25.92% in patients and 25.22% in controls. No significant differences were found between patients and controls in the frequency of HLA-B*51. Among kidney transplant recipients with HLA-B*51 (n = 72), 38.89% had rejection episodes and 61.11% had no rejection. No significant association was found between HLA-B*51 allele positivity and rejection. No significant relationship was shown between patients diagnosed with end-stage renal disease and HLA-B*51 positivity. Previous studies support frequency of the HLA-B*51 allele in the control group. Although Behçet disease is known to cause renal vasculitis, HLA-B*51 positivity alone was not associated with vasculitis or inflammation.

Protocol for Testing Human Melanoma Exosomes that Shift the Healthy Phenotype of Human Dermal Cells.

Exosomes are small membrane-derived vesicles that transmit DNA constituents, mRNAs, microRNAs, and proteins from donor cells to a receiver cell. Various cells comprising of mesenchymal, immune, and cancer cells discharge exosomes. Cancer cell exosomes form the entry and reprogramming of essentials connected to a tumor environment. Melanoma-derived exosomes transport diverse proteins such as c-MET and RAB27a, which leave a melanoma mark. Increased mesenchymal epithelial transition (MET) expressions in serum exosomes have been considered an indicator of disease progression. Meanwhile, RAB27a has been identified as being involved in exosome discharge and trafficking. Decreased expressions of RAB27a in human melanoma cells have shown to diminish exosome release.We examined the effects of the downregulation and upregulation of RAB27a and c-MET in human dermal fibroblasts by utilizing the isolated exosomes of malignant melanoma cell lines. Melanoma exosomes derived from cancer cells conveyed information to healthy dermal fibroblasts and stem cells while inducing phenotypic change. In this chapter, we show optimized protocols that were used by our group for in vitro analysis with melanoma exosomes.

The bone marrow of mouse-rat chimeras contains progenitors of multiple pulmonary cell lineages.

Radiation-induced lung injury (RILI) is a common complication of anti-cancer treatments for thoracic and hematologic malignancies. Bone marrow (BM) transplantation restores hematopoietic cell lineages in cancer patients. However, it is ineffective in improving lung repair after RILI due to the paucity of respiratory progenitors in BM transplants. In the present study, we used blastocyst injection to create mouse-rat chimeras, these are artificial animals in which BM is enriched with mouse-derived progenitor cells. FACS-sorted mouse BM cells from mouse-rat chimeras were transplanted into lethally irradiated syngeneic mice, and the contribution of donor cells to the lung tissue was examined using immunostaining and flow cytometry. Donor BM cells provided long-term contributions to all lung-resident hematopoietic cells which includes alveolar macrophages and dendritic cells. Surprisingly, donor BM cells also contributed up to 8% in pulmonary endothelial cells and stromal cells after RILI. To identify respiratory progenitors in donor BM, we performed single-cell RNA sequencing (scRNAseq). Compared to normal mouse BM, increased numbers of hematopoietic progenitors were found in the BM of mouse-rat chimeras. We also identified unique populations of hemangioblast-like progenitor cells expressing Hes1, Dntt and Ebf1, along with mesenchymal stromal cells expressing Cpox, Blvrb and Ermap that were absent or ultra-rare in the normal mouse BM. In summary, by using rats as "bioreactors", we created a unique mouse BM cell transplant that contributes to multiple respiratory cell types after RILI. Interspecies chimeras have promise for future generations of BM transplants enriched in respiratory progenitor cells.

Mitochondrial Transfer Between Mesenchymal Stem Cells and Cancer Cells.

Mitochondrial transfer (MT) is a biological process that allows a donor cell to horizontally share its own mitochondria with a recipient cell. Mitochondria are highly dynamic membrane-bound sub-cellular organelles prominently involved in the regulation of the cell energy balance, calcium homeostasis, and apoptotic machinery activation. They physiologically undergo fusion and fission processes in response to the cell requirement, with a continuous morphological re-arrangement. This structural and functional plasticity is at the basis of the MT, described in tissue regeneration, cardiac and neurological diseases, as well as in cancer. Here, the MT has been observed in the tumor micro-environment (TME) from the adipose-derived stem cells (ASCs) to the cancer cells, eventually reverting the lack of the mitochondria respiration function, or enhancing their motility and drug resistance. In this chapter, we outline some key protocols for evaluating this exciting phenomenon of MT. These methodological and technical approaches are very important, considering all the limitations that scientists constantly face, especially in this field of the research.