Distinct Stem Cell Populations Research Articles

Isolation and characterization of normal mandibular periosteal stem cells from human and macaca mulatta and cross-species single-cell analysis

Objective: To investigate the presence of a distinct stem cell populations different from mesenchymal stem cells in the mandibular periosteum of both human and non-human primates (macaca mulatta), to explore its properties during intramembranous osteogenesis and to establish standard protocols for the isolation, culturing and expanding of mandibular periosteal stem cells (PSC) distinguished from other PSCs in other anatomical regions. Methods: Periosteum was harvested from the bone surface during flap bone removal in patients aged 18-24 years undergoing third molar extraction and from the buccal side of the mandibular premolar region of 6-year-old macaca mulatta respectively, and then subjected to single-cell sequencing using the Illumina platform Novaseq 6000 sequencer. Cross-species single-cell transcriptome sequencing results were compared using homologous gene matching. PSC were isolated from primary tissues using two digestion methods with body temperature and low temperature, and their surface markers (CD200, CD31, CD45 and CD90) were identified by cell flow cytometry. The ability of cell proliferation and three-lineage differentiation of PSC expanded to the third generation in vitro in different species were evaluated. Finally, the similarities and differences in osteogenic properties of PSC and bone marrow mesenchymal stem cells (BMSC) were compared. Results: The single-cell sequencing results indicated that 18 clusters of cell populations were identified after homologous gene matching for dimensionality reduction, and manual cellular annotation was conducted for each cluster based on cell marker databases. The comparison of different digestion protocols proved that the low-temperature overnight digestion protocol can stably isolate PSC from the human and m. mulatta mandibular periosteum and the cells exhibited a fibroblast-like morphology. This research confirmed that PSC of human and m. mulatta had similar proliferation capabilities through the cell counting kit-8 assay. Flow cytometry analysis was then used to identify the cells isolated from the periosteum expressed CD200(+), CD31(-), CD45(-), CD90(-). Then, human and m. mulatta PSC were induced into osteogenesis, adipogenesis, and chondrogenesis to demonstrate their corresponding multi-lineage differentiation capabilities. Finally, comparison with BMSC further clarified the oesteogenesis characteristics of PSC. The above experiments proved that the cells isolated from the periosteum were peiosteal cells with characteristics of stem cells evidenced by their cell morphology, proliferation ability, surface markers, and differentiation ability, and that this group of PSC possessed characteristics different from traditional mesenchymal stem cells. Conclusions: In this study, normal mandibular PSC from humans and m. mulatta were stably isolated and identified for the first time, providing a cellular foundation for investigating the mechanism of mandibular intramembranous osteogenesis, exploring ideal non-human primate models and establishing innovative strategies for clinically mandibular injury repair.

Cell death as an architect of adult skin stem cell niches.

Our skin provides a physical and immunological barrier against dehydration and environmental insults ranging from microbial attacks, toxins and UV irradiation to wounding. Proper functioning of the skin barrier largely depends on the interplay between keratinocytes- the epithelial cells of the skin- and immune cells. Two spatially distinct populations of keratinocyte stem cells (SCs) maintain the epidermal barrier function and the hair follicle. These SCs are inherently long-lived, but cell death can occur within their niches and impacts their functionality. The default cell death programme in skin is apoptosis, an orderly and non-inflammatory suicide programme. However, recent findings are shedding light on the significance of various modes of regulated necrotic cell death, which are lytic and can provoke inflammation within the local skin environment. While the presence of dying cells was generally regarded as a mere consequence of inflammation, findings in various human dermatological conditions and experimental mouse models of aberrant cell death control demonstrated that cell death programmes in keratinocytes (KCs) can drive skin inflammation and even tumour initiation. When cells die, they need to be removed by phagocytosis and KCs can function as non-professional phagocytes of apoptotic cells with important implications for their SC capacities. It is becoming apparent that in conditions of heightened SC activity, distinct cell death modalities differentially impact the different skin SC populations in their local niches. Here, we describe how regulated cell death modalities functionally affect epidermal SC niches along with their relevance to injury repair, inflammatory skin disorders and cancer.

Lepr-Expressing PDLSCs Contribute to Periodontal Homeostasis and Respond to Mechanical Force by Piezo1.

Periodontium supports teeth in a mechanically stimulated tissue environment, where heterogenous stem/progenitor populations contribute to periodontal homeostasis. In this study, Leptin receptor+ (Lepr+) cells are identified as a distinct periodontal ligament stem cell (PDLSC) population by single-cell RNA sequencing and lineage tracing. These Lepr+ PDLSCs are located in the peri-vascular niche, possessing multilineage potential and contributing to tissue repair in response to injury. Ablation of Lepr+ PDLSCs disrupts periodontal homeostasis. Hyper-loading and unloading of occlusal forces modulate Lepr+ PDLSCs activation. Piezo1 is demonstrated that mediates the mechanosensing of Lepr+ PDLSCs by conditional Piezo1-deficient mice. Meanwhile, Yoda1, a selective activator of Piezo1, significantly accelerates periodontal tissue growth via the induction of Lepr+ cells. In summary, Lepr marks a unique multipotent PDLSC population in vivo, to contribute toward periodontal homeostasis via Piezo1-mediated mechanosensing.

2023 TERMIS – AMERICAS Conference & Exhibition Boston Marriott Copley Place April 11–14, 2023

2023 TERMIS – AMERICAS Conference & Exhibition Boston Marriott Copley Place April 11–14, 2023

Progress and challenges in stem cell biology.

Since stem cells were first discovered, researchers have identified distinct stem cell populations in different organs and with various functions, converging on the unique abilities of self-renewal and differentiation toward diverse cell types. These abilities make stem cells an incredibly promising tool in therapeutics and have turned stem cell biology into a fast-evolving field. Here, stem cell biologists express their view on the most striking advances and current challenges in their field.

MECHANISMS OF DEVELOPMENT AND REGENERATION IN HYDRA

Abstract Hydra vulgaris is a small and simple aquatic animal capable of whole-body regeneration and has negligible senescence. The entire animal, including the nervous system, is composed of about 25 cell types, and can regenerate from a fragment of tissue as small as ~300 cells. In addition, all cell types are continually renewed in the uninjured adult as part of normal homeostasis; every differentiated cell type is replaced approximately every 20 days, which likely contributes to its lack of aging. The remarkable features of Hydra are enabled by three distinct populations of stem cells that support the three lineages that make up the adult Hydra – the ectodermal epithelial lineage, the endodermal epithelial lineage, and the interstitial lineage (includes the neurons). A major goal of our laboratory is to understand the gene regulatory networks that control the specification of all Hydra cell types in the uninjured (homeostatic) state and then understand how injury triggers these differentiation pathways at unexpected locations during regeneration. Using high throughput genomics approaches such as scRNA-seq, ATAC-seq, and Cut&Tag, we have transcriptionally defined every cell type in Hydra and identified putative transcriptional regulators for each cell type. This includes the 11 neuronal subtypes that comprise the nerve net that spans the entire length of the Hydra body. We are currently leveraging these data to conduct functional testing of key putative regulators and to identify injury inputs into cell specification events during regeneration.

Distinct bulge stem cell populations maintain the pilosebaceous unit in a β-catenin-dependent manner

The pilosebaceous unit (PSU) is composed of multiple compartments and the self-renewal of PSU depends on distinct hair follicle stem cell (HFSC) populations. However, the differential roles of the HFSCs in sebaceous gland (SG) renewal have not been completely understood. Here, we performed multiple lineage tracing analysis to unveil the contribution of different HFSC populations to PSU regeneration during the hair cycle and wound healing. Our results indicated that the upper bulge stem cells contributed extensively to the SG replenishment during hair cycling, while HFSCs in the lower bugle did not. During skin wound healing, all HFSC populations participated in the SG replenishment. Moreover, β-catenin activation promoted the contribution of HFSCs to SG replenishment, whereas β-catenin deletion substantially repressed the event. Thus, our findings indicated that HFSCs contributed to SG replenishment in a β-catenin-dependent manner.

Isolation of Quiescent Stem Cell Populations from Individual Skeletal Muscles.

Skeletal muscle harbors distinct populations of adult stem cells that contribute to the homeostasis and repair of the tissue. Skeletal muscle stem cells (MuSCs) have the ability to make new muscle, whereas fibro-adipogenic progenitors (FAPs) contribute to stromal supporting tissues and have the ability to make fibroblasts and adipocytes. Both MuSCs and FAPs reside in a state of prolonged reversible cell cycle exit, called quiescence. The quiescent state is key to their function. Quiescent stem cells are commonly purified from multiple muscle tissues pooled together in a single sample. However, recent studies have revealed distinct differences in the molecular profiles and quiescence depth of MuSCs isolated from different muscles. The present protocol describes the isolation and study of MuSCs and FAPs from individual skeletal muscles and presents strategies to perform molecular analysis of stem cell activation. It details how to isolate and digest muscles of different developmental origin, thicknesses, and functions, such as the diaphragm, triceps, gracilis, tibialis anterior (TA), gastrocnemius (GA), soleus, extensor digitorum longus (EDL), and the masseter muscles. MuSCs and FAPs are purified by fluorescence-activated cell sorting (FACS) and analyzed by immunofluorescence staining and 5-ethynyl-2´-deoxyuridine (EdU) incorporation assay.

Single-Cell Multiomics of Pre- and Post- Remission Bone Marrow from a Pediatric Patient with JMML Reveal Distinct Progenitors and Inflammatory Monocytes Which Are Eliminated By Chemotherapy

Juvenile myelomonocytic leukemia (JMML) is a rare myeloproliferative/myelodysplastic disorder with a poor prognosis. Unlike many other hematologic malignancies, it remains challenging to rapidly diagnose, and levels of residual disease are challenging to measure. Ongoing efforts to characterize the pathophysiology of JMML have yielded significant insight, but a complete picture of the cellular hierarchy of the disease has been elusive. To obtain an unsupervised view of the heterogeneity of hematopoietic cell types in JMML, we performed single-cell RNA multiomics on serial samples from a 4-year-old patient who presented with fevers, hepatosplenomegaly, leukocytosis, monocytosis, elevated Hemoglobin F, and bone marrow blast count of 7%. Clinical sequencing revealed mutations in PTPN11 (VAF 66%), NF1 (VAF 17%), MYB (VAF 12%), and SH2B3. We obtained a sample for multiomic profiling following his initial cycles of chemotherapy when PTPN11 variant frequency was 32%, and he remained critically ill. The subsequent sample (6 weeks later) was collected when no mutated PTPN11 was detectible, and his clinical picture had significantly improved. Both samples were processed using gradient centrifugation, and a subset of the cells captured was immunomagnetically enriched CD34+ cells. A panel of 141 oligonucleotide-tagged antibodies was used to label the cells prior to single-cell capture. The resulting libraries were sequenced using standard short-read sequencing, and reads were demultiplexed and counted using common workflows. Single-cell profiles were visualized using UMAP. Cells were annotated using a highly performant machine-learning algorithm that classifies cells using a healthy reference of human BM, which we have previously validated using publicly available data. Examining all cells together, we observed changes in two key cell types over time (Figure 1). First, we noted a large population of monocyte-like cells with increased expression of genes associated with complement, inflammation (including IL1beta), and KRAS signaling in the pre-remission sample, which nearly completely disappeared in the second sample. Moreover, surface expression of CD39 was significantly higher in these cells than in all other cells measured (Figure 2). These data suggest CD39 may represent a novel biomarker to monitor treatment response in JMML or as a potential target for therapy. Second, we observed two distinct hematopoietic stem cell (HSC) populations in the pre-remission sample and only one in the remission sample. Differential gene expression analysis between the two pre-remission HSC clusters revealed enrichment of specific genes previously associated with JMML progenitors in one, including HOPX. This population of HSCs also exhibited higher expression CD7, which correlated with the patient's flow cytometric data from the same time point. We hypothesize that CD7/HOPX expressing HSCs in the first sample were JMML progenitor cells eliminated by therapy. Our work demonstrates the power of single-cell multiomics to dissect cellular heterogeneity in diseases with complex clonal hierarchies. Moreover, these data can also be a resource for identifying potential biomarkers and drug targets. Specifically, our data suggest that inflammatory monocytes may be a distinct target for therapy, which may benefit critically ill patients like ours. Future efforts will be focused on profiling additional patients and rigorously characterizing the mutational state of individual cells. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Plasticity of Epithelial Cells during Skin Wound Healing.

Epithelial tissues line the outer surfaces of the mammalian body and protect from external harm. In skin, the epithelium is maintained by distinct stem cell populations residing in the interfollicular epidermis and various niches of the hair follicle. These stem cells give rise to the stratified epidermal layers and the protective hair coat, while being confined to their respective niches. Upon injury, however, all stem cell progenies can leave their niche and collectively contribute to a central wound healing process, called reepithelialization, for restoring the skin's barrier function. This review explores how epithelial cells from distinct niches respond and adapt during acute wound repair. We discuss when and where cells sense and react to damage, how cellular identity is regulated at the molecular and behavioral level, and how cells memorize past experiences and their origin. This collective knowledge highlights cellular plasticity as a brilliant feature of epithelial tissues to heal.

Baicalin regulates stem cells as a creative point in the treatment of climacteric syndrome.

It is widely acknowledged that the climacteric syndrome negatively affects women's quality of life and leads to cerebral ischemic injury, osteoporosis and cardiovascular disease. One of the main active ingredients in Radix Scutellariae, Baicalin, has been established to possess a wide range of pharmacological effects and is beneficial in enhancing osteogenic differentiation and cardiovascular disease. Baicalin's profound metabolic impact on various stem cell populations and their fate specification could improve the efficiency of stem cell therapy for climacteric syndrome. However, Baicalin-mediated processes are complex and many of the underlying mechanisms are not fully fathomed yet. This review aims to shed light on the regulatory role of Baicalin on the diverse behaviors of distinct stem cell populations and provide a good cell source for stem cell therapy to broaden the therapeutic landscape for climacteric syndrome patients.

Promises and Challenges of Cell-Based Therapies to Promote Lung Regeneration in Idiopathic Pulmonary Fibrosis.

The lung epithelium is constantly exposed to harmful agents present in the air that we breathe making it highly susceptible to damage. However, in instances of injury to the lung, it exhibits a remarkable capacity to regenerate injured tissue thanks to the presence of distinct stem and progenitor cell populations along the airway and alveolar epithelium. Mechanisms of repair are affected in chronic lung diseases such as idiopathic pulmonary fibrosis (IPF), a progressive life-threatening disorder characterized by the loss of alveolar structures, wherein excessive deposition of extracellular matrix components cause the distortion of tissue architecture that limits lung function and impairs tissue repair. Here, we review the most recent findings of a study of epithelial cells with progenitor behavior that contribute to tissue repair as well as the mechanisms involved in mouse and human lung regeneration. In addition, we describe therapeutic strategies to promote or induce lung regeneration and the cell-based strategies tested in clinical trials for the treatment of IPF. Finally, we discuss the challenges, concerns and limitations of applying these therapies of cell transplantation in IPF patients. Further research is still required to develop successful strategies focused on cell-based therapies to promote lung regeneration to restore lung architecture and function.

Thy1 marks a distinct population of slow-cycling stem cells in the mouse epidermis

The presence of distinct stem cells that maintain the interfollicular epidermis is highly debated. Here, we report a population of keratinocytes, marked by Thy1, in the basal layer of the interfollicular epidermis. We find that epidermal cells expressing differential levels of Thy1 display distinct transcriptional signatures. Thy1+ keratinocytes do not express T cell markers, express a unique transcriptional profile, cycle significantly slower than basal epidermal progenitors and display significant expansion potential in vitro. Multicolor lineage tracing analyses and mathematical modeling reveal that Thy1+ basal keratinocytes do not compete neutrally alike interfollicular progenitors and contribute long-term to both epidermal replenishment and wound repair. Importantly, ablation of Thy1+ cells strongly impairs these processes, thus indicating the non-redundant function of Thy1+ stem cells in the epidermis. Collectively, these results reveal a distinct stem cell population that plays a critical role in epidermal homeostasis and repair.

Uncovering the spectrum of adult zebrafish neural stem cell cycle regulators.

Adult neural stem and progenitor cells (aNSPCs) persist lifelong in teleost models in diverse stem cell niches of the brain and spinal cord. Fish maintain developmental stem cell populations throughout life, including both neuro-epithelial cells (NECs) and radial-glial cells (RGCs). Within stem cell domains of the brain, RGCs persist in a cycling or quiescent state, whereas NECs continuously divide. Heterogeneous populations of RGCs also sit adjacent the central canal of the spinal cord, showing infrequent proliferative activity under homeostasis. With the rise of the zebrafish (Danio rerio) model to study adult neurogenesis and neuroregeneration in the central nervous system (CNS), it has become evident that aNSPC proliferation is regulated by a wealth of stimuli that may be coupled with biological function. Growing evidence suggests that aNSPCs are sensitive to environmental cues, social interactions, nutrient availability, and neurotrauma for example, and that distinct stem and progenitor cell populations alter their cell cycle activity accordingly. Such stimuli appear to act as triggers to either turn on normally dormant aNSPCs or modulate constitutive rates of niche-specific cell cycle behaviour. Defining the various forms of stimuli that influence RGC and NEC proliferation, and identifying the molecular regulators responsible, will strengthen our understanding of the connection between aNSPC activity and their biological significance. In this review, we aim to bring together the current state of knowledge on aNSPCs from studies investigating the zebrafish CNS, while highlighting emerging cell cycle regulators and outstanding questions that will help to advance this fascinating field of stem cell biology.

29. MECHANICAL STRETCH MOBILIZES LGR6+ EPIDERMAL STEM CELLS TO DRIVE SKIN GROWTH

PURPOSE: To uncover cellular and molecular determinants of skin growth with the goal of designing future therapies for conditions of excess or inadequate skin. Current research in the field has identified distinct populations of stem cells within specific skin components that regularly replenish the epidermis and hair follicles. METHODS: A controlled tissue expansion mouse model was applied to all variables of interest. We used Lgr5/6-EGFP-CreERT2;tdTomato mice to lineage trace Lgr5/6+ stem cell progeny. YAP gain- and loss-of-function mouse models quantified YAP expression. 3D skin model reconstructions characterized skin components. RNA microarray and single-cell RNA sequencing identified changes in metabolic pathways. Pseudotime analysis assessed lineage trajectory of Lgr6+ cells. RESULTS: Skin growth from mechanical tension is driven by proliferation of the epidermis. Epidermal growth is achieved through preferential activation and differentiation of not Lgr5+, but instead Lgr6+ stem cells of the interfollicular epidermis, driven in part by activation of the Hippo pathway effector YAP. A number of growth regulatory (PI3K-AKT), inflammatory (Toll-like receptor), and oxygen sensing (Hif1) pathways were up-regulated. CONCLUSION: By defining the differential responses of skin elements to tension-dependent growth, we begin to identify targets to improve skin generation. Methods that may lead to more effective expansion include promoting fibroblast expansion, increasing the number of Lgr6+ stem cells, and direct activation of tension-responsive elements such as YAP. Up-regulated pathways offer additional molecular targets for enhancing skin growth. Future research may investigate the role of mechanical signaling as a driver of growth in other organs such as gut and lung.

3193 – EXPRESSION OF CD61 MARKS FUNCTIONALLY DISTINCT POPULATIONS OF AGED HEMATOPOIETIC STEM CELLS

Hematopoietic stem cells (HSCs) give rise to all hematopoietic cells throughout the lifespan of an organism. Upon ageing, murine HSCs gradually lose their self-renewal and regenerative potential, skew towards the myeloid lineage and expand in numbers. Also, the aged HSC pool becomes increasingly heterogeneous where some cells have remained ‘young-like'. Here we aimed to identify and isolate these ‘young-like' HSCs from aged mice to better understand the mechanisms that contribute to HSC ageing. We focused on CD61 (Integrin beta 3), a membrane-associated gene that we previously identified as one of the most consistently differentially expressed gene between young and old murine HSCs. CD61 plays a role in a variety of cellular processes, including cell adhesion and migration. We hypothesized that the level of CD61 expression on HSCs marks distinct populations of stem cells. We competitively transplanted 500 CD61High and 500 CD61Low young and aged long-term hematopoietic stem cells (LT-HSCs) alongside 2x106 competitor bone marrow cells into lethally irradiated recipients. Chimerism levels were analyzed by FACS every 4 weeks for 20 weeks. LT-HSCs from primary recipients were then competitively transplanted into secondary recipients to further investigate their functionality. In primary transplantation, young CD61High and CD61Low HSCs provided comparable engraftment. However, CD61High HSCs from aged donors resulted in, on average, 80% chimerism whereas the CD61Low HSCs provided only 20% chimerism. In contrast to primary transplantations, secondary transplantations with young CD61Low HSCs resulted in 20% lower chimerism compared to CD61High HSCs. Our data demonstrate that expression of CD61 becomes increasingly important as HSCs age. In addition, our data from secondary transplantation suggest that CD61 might mark intrinsically superior cells in the HSCs pool.

SATB2 preserves colon stem cell identity and mediates ileum-colon conversion via enhancer remodeling.

Adult stem cells maintain regenerative tissue structure and function by producing tissue-specific progeny, but the factors that preserve their tissue identities are not well understood. The small and large intestines differ markedly in cell composition and function, reflecting their distinct stem cell populations. Here we show that SATB2, a colon-restricted chromatin factor, singularly preserves LGR5+ adult colonic stem cell and epithelial identity in mice and humans. Satb2 loss in adult mice leads to stable conversion of colonic stem cells into small intestine ileal-like stem cells and replacement of the colonic mucosa with one that resembles the ileum. Conversely, SATB2 confers colonic properties on the mouse ileum. Human colonic organoids also adopt ileal characteristics upon SATB2 loss. SATB2 regulates colonic identity in part by modulating enhancer binding of the intestinal transcription factors CDX2 and HNF4A. Our study uncovers a conserved core regulator of colonic stem cells able to mediate cross-tissue plasticity in mature intestines.

Airbrushed nanofibrous membranes to control stem cell infiltration in 3D‐printed scaffolds

AbstractExtrusion‐based 3D printing of polymeric biomaterials has emerged as a promising approach for the fabrication of complex tissue engineering constructs. However, the large pore and feature size lead to low cell seeding efficiency and limited control of spatial distribution of cells within the scaffolds. We developed hybrid scaffolds that are composed of 3D‐printed layers and airbrushed fibrous membranes. Airbrushing time was adjusted to fabricate low (L), medium (M), and high (H) density membranes to effectively control stem cell infiltration. When two distinct populations of stem cells were seeded from top or bottom of the scaffolds, scaffolds composed of LLL membranes showed gradual mixing of the cells with depth, whereas LHL membranes led to two distinct regions of cells separated by the H membrane. Our results demonstrate that fibrous membranes incorporated within 3D‐printed layers enable user‐defined and spatially controlled cell compositions within hybrid scaffolds.

Immunohistochemical identification of complement peptide C5a receptor 1 (C5aR1) in non-neoplastic and neoplastic human tissues.

The complement component C5a and its receptor C5aR1 are involved in the development of numerous inflammatory diseases. In addition to immune cells, C5aR1 is expressed in neoplastic cells of multiple tumour entities, where C5aR1 is associated with a higher proliferation rate, advanced tumour stage, and poor patient outcomes. The aim of the present study was to obtain a broad expression profile of C5aR1 in human non-neoplastic and neoplastic tissues, especially in tumour entities not investigated in this respect so far. For this purpose, we generated a novel polyclonal rabbit antibody, {5227}, against the carboxy-terminal tail of C5aR1. The antibody was initially characterised in Western blot analyses and immunocytochemistry using transfected human embryonic kidney (HEK) 293 cells. It was then applied to a large series of formalin-fixed, paraffin-embedded non-neoplastic and neoplastic human tissue samples. C5aR1 was strongly expressed by different types of immune cells in the majority of tissue samples investigated. C5aR1 was also present in alveolar macrophages, bronchial, gut, and bile duct epithelia, Kupffer cells, occasionally in hepatocytes, proximal renal tubule cells, placental syncytiotrophoblasts, and distinct stem cell populations of bone marrow. C5aR1 was also highly expressed in the vast majority of the 32 tumour entities investigated, where a hitherto unappreciated high prevalence of the receptor was detected in thyroid carcinomas, small-cell lung cancer, gastrointestinal stromal tumours, and endometrial carcinomas. In addition to confirming published findings, we found noticeable C5aR1 expression in many tumour entities for the first time. Here, it may serve as an interesting target for future therapies.

Mitochondrial metabolism is a key regulator of the fibro-inflammatory and adipogenic stromal subpopulations in white adipose tissue.

The adipose tissue stroma is a rich source of molecularly distinct stem and progenitor cell populations with diverse functions in metabolic regulation, adipogenesis, and inflammation. The ontology of these populations and the mechanisms that govern their behaviors in response to stimuli, such as overfeeding, however, are unclear. Here, we show that the developmental fates and functional properties of adipose platelet-derived growth factor receptor beta (PDGFRβ)+ progenitor subpopulations are tightly regulated by mitochondrial metabolism. Reducing the mitochondrial β-oxidative capacity of PDGFRβ+ cells via inducible expression of MitoNEET drives a pro-inflammatory phenotype in adipose progenitors and alters lineage commitment. Furthermore, disrupting mitochondrial function in PDGFRβ+ cells rapidly induces alterations in immune cell composition in lean mice and impacts expansion of adipose tissue in diet-induced obesity. The adverse effects on adipose tissue remodeling can be reversed by restoring mitochondrial activity in progenitors, suggesting therapeutic potential for targeting energy metabolism in these cells.