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Article
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A microfluidic platform integrating functional vascularized organoids-on-chip
Vascularization remains a significant challenge in organoid technology. Here, the authors develop a microfluidic platform that enhances organoid growth, function and maturation, by establishing functional perfusable vascular networks.
- Clément Quintard
- , Emily Tubbs
- & Xavier Gidrol
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Chemically-defined and scalable culture system for intestinal stem cells derived from human intestinal organoids
Challenges in reproducibility and large-scale expansion limit the current applicability of human intestinal organoids. Here, the authors present a feeder-free, chemically-defined culture method for enrichment of intestinal stem cells isolated from 3D human intestinal organoids.
- Ohman Kwon
- , Hana Lee
- & Mi-Young Son
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Generation of three-dimensional meat-like tissue from stable pig epiblast stem cells
Cultured meat technology promises to alleviate protein shortages, but still faces many challenges. Here, the authors achieve serum-free myogenic differentiation of porcine pre-gastrulation epiblast stem cells and generate meat-like tissue via edible plant-based scaffolds without any animal compounds.
- Gaoxiang Zhu
- , Dengfeng Gao
- & Jianyong Han
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Enhanced pericyte-endothelial interactions through NO-boosted extracellular vesicles drive revascularization in a mouse model of ischemic injury
Treatment alternatives are not available for a significant portion of critical limb ischemia patients. Here, the authors show a strategy to engineer mesenchymal stem cells-derived extracellular vesicles to induce revascularization in mice.
- Ling Guo
- , Qiang Yang
- & Min Feng
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Mass production of lumenogenic human embryoid bodies and functional cardiospheres using in-air-generated microcapsules
Current methods to generate spheroids are associated with low production throughputs, limiting clinical and industrial translation. Here the authors present a clean ultra-high-throughput in-air microfluidic platform for mass production of lumenogenic embryoid bodies and functional cardiospheres.
- Bas van Loo
- , Simone A. ten Den
- & Jeroen Leijten
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Mechanically enhanced biogenesis of gut spheroids with instability-driven morphomechanics
Understanding and controlling morphogenesis is vital for biology and organoid technology. Here, the authors report an efficient biomechanical system to generate gut spheroids, and reveal instability-driven morphogenetic transitions with computational models.
- Feng Lin
- , Xia Li
- & Yue Shao
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A village in a dish model system for population-scale hiPSC studies
Village cultures, where multiple stem cell lines are cultured in a single dish, provide an elegant solution for population-scale studies. Here, authors show the utility of village models – showing that expression heterogeneity is largely a result of line-specific effects and not village cultures.
- Drew R. Neavin
- , Angela M. Steinmann
- & Joseph E. Powell
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Hematopoietic reconstitution dynamics of mobilized- and bone marrow-derived human hematopoietic stem cells after gene therapy
Scala et al. show that mobilized peripheral blood hematopoietic stem/progenitor cells are more enriched in repopulating stem cells than bone marrow. Moreover, the quantity and type of infused subsets correlated with gene therapy outcome in humans.
- Serena Scala
- , Francesca Ferrua
- & Alessandro Aiuti
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CAR-neutrophil mediated delivery of tumor-microenvironment responsive nanodrugs for glioblastoma chemo-immunotherapy
Neutrophil-mediated drug delivery has been investigated as a therapeutic approach for brain tumors. Here the authors report the anti-tumor activity of chlorotoxin-directed CAR neutrophils delivering chemodrug-loaded nanoparticles in preclinical glioblastoma models.
- Yun Chang
- , Xuechao Cai
- & Xiaoping Bao
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Expansion of human megakaryocyte-biased hematopoietic stem cells by biomimetic Microniche
An effective expansion system for therapeutic megakaryocyte-biased hematopoietic stem cells has not been developed. Here the authors present a microniche-based system that supports scalable expansion of human Mk-biased HSCs and identify their immune phenotype.
- Yinghui Li
- , Mei He
- & Yingdai Gao
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Retinoic acid signaling modulation guides in vitro specification of human heart field-specific progenitor pools
The heart is formed from several spatiotemporally distinct progenitor pools during development. Here they show that modulation of retinoic acid signaling can instruct human pluripotent stems cells into heart progenitors that are useful for studying human development and disease.
- Dorota Zawada
- , Jessica Kornherr
- & Alexander Goedel
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Therapeutic adenine base editing of human hematopoietic stem cells
Here, Liao and colleagues apply adenine base editor ABE8e and its PAM-less variant ABE8e-SpRY to β-thalassemia patient hematopoietic stem cells in the form of ribonucleoprotein complexes, resulting in efficient long-term editing and β-thalassemia alleviation.
- Jiaoyang Liao
- , Shuanghong Chen
- & Yuxuan Wu
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Retrotransposon instability dominates the acquired mutation landscape of mouse induced pluripotent stem cells
Retrotransposons are mobile genetic elements normally repressed by DNA methylation in differentiated cells. Here, the authors show that DNA hypomethylation in mouse induced pluripotent stem cells allows retrotransposons to jump, but this can be blocked with a reverse transcriptase inhibitor.
- Patricia Gerdes
- , Sue Mei Lim
- & Geoffrey J. Faulkner
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Bead-jet printing enabled sparse mesenchymal stem cell patterning augments skeletal muscle and hair follicle regeneration
Current mesenchymal stem cell (MSC) transplantation practices are limited by the loss or reduced performance of MSCs. Here the authors develop a bead-jet printer for intraoperative formulation and printing of MSCs-laden Matrigel beads to improve skeletal muscle and hair follicle regeneration.
- Yuanxiong Cao
- , Jiayi Tan
- & Shaohua Ma
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Transplantable human thyroid organoids generated from embryonic stem cells to rescue hypothyroidism
Hypothyroidism is a common condition that cannot always be satisfactorily treated through continued hormone replacement. Here the authors report the generation of transplantable thyroid organoids derived from human embryonic stem cells that can restore plasma thyroid hormone in athyreotic mice.
- Mírian Romitti
- , Adrien Tourneur
- & Sabine Costagliola
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CHD7 regulates otic lineage specification and hair cell differentiation in human inner ear organoids
Mutations in the chromatin remodeler CHD7 cause CHARGE syndrome, affecting development of several organs including the inner ear. Here, the authors recapitulated pathogenesis of this disease with human inner ear organoids and found that CHD7 is indispensable for proper otic lineage specification and hair cell differentiation.
- Jing Nie
- , Yoshitomo Ueda
- & Eri Hashino
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Nanozyme-reinforced hydrogel as a H2O2-driven oxygenerator for enhancing prosthetic interface osseointegration in rheumatoid arthritis therapy
Stem cell transplantation has huge potential but can suffer from effects caused by disease state microenvironment. Here, the authors report on the development of a nanozyme reinforced hydrogel for reactive oxygen species scavenging and the generation of dissolved oxygen, demonstrating improved osteointegration using the hydrogel.
- Yue Zhao
- , Shanliang Song
- & Yanli Zhao
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Transient inhibition of p53 enhances prime editing and cytosine base-editing efficiencies in human pluripotent stem cells
Li et al. report that co-delivering a dominant negative fragment of p53 (p53DD) greatly enhances precise editing efficiencies for prime editing and cytosine base editing in human pluripotent stem cells, without compromising the genome-wide safety.
- Mu Li
- , Aaron Zhong
- & Ting Zhou
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Salivary gland organoid culture maintains distinct glandular properties of murine and human major salivary glands
The long-term maintenance of diverse salivary gland cells remains challenging. Here the authors establish a protocol for long-term maintenance of murine and human salivary gland organoids exhibiting gland-specific gene expression, gland functions, and cellular diversity confirmed by scRNA-seq.
- Yeo-Jun Yoon
- , Donghyun Kim
- & Jae-Yol Lim
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Open-source personal pipetting robots with live-cell incubation and microscopy compatibility
Liquid handling and pipetting tools can automate repetitive tasks but are far from universally used. Here the authors report the Pipetting Helper Imaging Lid (PHIL), an open-source liquid handling robot designed for inexperienced users, that they use for automated pipetting.
- Philip Dettinger
- , Tobias Kull
- & Timm Schroeder
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Signal requirement for cortical potential of transplantable human neuroepithelial stem cells
The regulatory pathways that control the human neural progenitor cell pool are not well understood. Here, Varga et al. identify signals that control the division of human pluripotent stem cell derived neural stem cells and their ability to make cortical neurons and glia.
- Balazs V. Varga
- , Maryam Faiz
- & Andras Nagy
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Tissue extracellular matrix hydrogels as alternatives to Matrigel for culturing gastrointestinal organoids
The culture of gastrointestinal organoids relies on Matrigel that has several drawbacks for clinical application. Here, the authors report the feasibility of gastrointestinal tissue-mimetic matrices as effective alternatives to Matrigel for organoid culture and transplantation.
- Suran Kim
- , Sungjin Min
- & Seung-Woo Cho
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Self-organization of human dorsal-ventral forebrain structures by light induced SHH
Organizing centers act to pattern surrounding tissues during embryogenesis through the secretion of morphogens. Here the authors model human organizers using light stimulus to geometrically confine SHH expression in differentiating hESCs, generating spatially resolved proximal distal patterns.
- Riccardo De Santis
- , Fred Etoc
- & Ali H. Brivanlou
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StemBond hydrogels control the mechanical microenvironment for pluripotent stem cells
The independent control of substrate stiffness and tethering of extracellular matrix to substrates for mechanical signalling investigations remains challenging. Here the authors present StemBond hydrogels, with stable ECM tethering that can be varied independently of stiffness, and use these to modulate the function of mouse and human pluripotent stem cells.
- Céline Labouesse
- , Bao Xiu Tan
- & Kevin J. Chalut
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A CRISPR/Cas9 genetically engineered organoid biobank reveals essential host factors for coronaviruses
Rapid identification of host genes essential for virus replication may expedite the generation of therapeutic interventions. Here the authors generate mutant clonal intestinal organoids for 19 host genes previously implicated in coronavirus biology and identify the cell surface protease TMPRSS2 as a potential therapeutic target.
- Joep Beumer
- , Maarten H. Geurts
- & Hans Clevers
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Improved modeling of human AD with an automated culturing platform for iPSC neurons, astrocytes and microglia
Human induced pluripotent stem cell (iPSC) cells have been used to model disease in specific cell types. Here, the authors develop an automated long-term culturing platform of human iPSC neurons, astrocytes, and microglia and use it to model some cellular aspects of Alzheimer’s disease.
- Reina Bassil
- , Kenneth Shields
- & Ben Chih
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Engineered whole cut meat-like tissue by the assembly of cell fibers using tendon-gel integrated bioprinting
Mammalian cell-based cultured meat has mostly been unstructured, leaving a demand for artificial steak-like meat. Here the authors present an assembled steak-like tissue of bovine skeletal muscle, adipose tissue, and blood capillary tissue fabricated by tendon-gel integrated printing technology.
- Dong-Hee Kang
- , Fiona Louis
- & Michiya Matsusaki
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Microfluidic device with brain extracellular matrix promotes structural and functional maturation of human brain organoids
Brain organoids derived from human pluripotent stem cells can model human brain development and disease, though current culture systems fail to ensure reliable production of high-quality organoids. Here the authors combine human brain extracellular matrix and culture in a microfluidic device to promote structural and functional maturation of human brain organoids.
- Ann-Na Cho
- , Yoonhee Jin
- & Seung-Woo Cho
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Label-free imaging for quality control of cardiomyocyte differentiation
Differentiation of hPSCs to cardiomyocytes suffers from high variability. Here the authors report a label-free live cell imaging platform based on autofluorescence imaging to enable the prediction of cardiomyocyte differentiation efficiency from hPSCs.
- Tongcheng Qian
- , Tiffany M. Heaster
- & Melissa C. Skala
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Establishment of a fluorescent reporter of RNA-polymerase II activity to identify dormant cells
The identification and characterisation of dormant cells is currently difficult. Here the authors report Optical Stem Cell Activity Reporter (OSCAR) to assess RNA polymerase II activity and identify dormant cell populations in intestinal epithelial cells in vivo.
- Rasmus Freter
- , Paola Falletta
- & Francesco Neri
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The TRACE-Seq method tracks recombination alleles and identifies clonal reconstitution dynamics of gene targeted human hematopoietic stem cells
Genetic barcoding has been used to track clonal dynamics of cells. Here, the authors develop a Tracking Recombination Alleles in Clonal Engraftment using sequencing (TRACE-Seq), to barcode repaired alleles by introducing silent mutations or outside of coding regions, to show clonal complexity of edited CD34 + cells following engraftment.
- Rajiv Sharma
- , Daniel P. Dever
- & Ravindra Majeti
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Automated microfluidic platform for dynamic and combinatorial drug screening of tumor organoids
The use of organoids in personalized medicine is promising but high throughput platforms are needed. Here the authors develop an automated, high-throughput, microfluidic 3D organoid culture system that allows combinatorial and dynamic drug treatments and real-time analysis of organoids.
- Brooke Schuster
- , Michael Junkin
- & Savaş Tay
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Vulnerability of progeroid smooth muscle cells to biomechanical forces is mediated by MMP13
Hutchinson-Gilford Progeria Syndrome (HGPS) is a premature aging disease and smooth muscle cells are the most affected cells in HGPS individuals. Here, the authors report a microfluidics platform with HGPS induced pluripotent stem cells and show that inhibition of metalloprotease 13 may reduce smooth muscle cell loss.
- Patricia R. Pitrez
- , Luís Estronca
- & Lino Ferreira
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Targeted gene correction of human hematopoietic stem cells for the treatment of Wiskott - Aldrich Syndrome
In recent years, hematopoietic stem cells gene editing has emerged as a promising tool to treat blood disorders. Here the authors develop a CRISPR/Cas9-based genome editing strategy that allows the precise correction of Wiskott-Aldrich Syndrome in vitro and in vivo with high efficiency.
- Rajeev Rai
- , Marianna Romito
- & Alessia Cavazza
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Stem cell-derived polarized hepatocytes
To model hepatocyte function accurately in vitro, it is necessary to generate and maintain a polarized epithelium. Here, the authors describe a protocol to generate polarized human pluripotent stem cell-derived hepatocyte-like cells (HLCs) to model enteric virus production and drug secretion in vitro.
- Viet Loan Dao Thi
- , Xianfang Wu
- & Charles M. Rice
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Transient non-integrative expression of nuclear reprogramming factors promotes multifaceted amelioration of aging in human cells
Aging involves gradual loss of tissue function, and transcription factor (TF) expression can ameliorate this in progeroid mice. Here the authors show that transient TF expression reverses age-associated epigenetic marks, inflammatory profiles and restores regenerative potential in naturally aged human cells.
- Tapash Jay Sarkar
- , Marco Quarta
- & Vittorio Sebastiano
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Caliciviral protein-based artificial translational activator for mammalian gene circuits with RNA-only delivery
Synthetic RNA-based circuits allow for gene regulation without the risk of mutagenesis. Here the authors demonstrate a Caliciviral VPg-based Translational activator which allows translational activation of synthetic mRNAs.
- Hideyuki Nakanishi
- & Hirohide Saito
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Modeling medulloblastoma in vivo and with human cerebellar organoids
Group 3 medulloblastoma (MB) is considered one of the most aggressive forms of this cancer. Here, the authors show that Otx2 and c-MYC oncogenes can drive Group 3 MB formation in mouse and human cerebellar organoids while SMARCA4 overexpression or a EZH2-specific inhibitor can inhibit tumorigenesis.
- Claudio Ballabio
- , Marica Anderle
- & Luca Tiberi
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Generating ring-shaped engineered heart tissues from ventricular and atrial human pluripotent stem cell-derived cardiomyocytes
The cellular composition of previous engineered heart tissue is often heterogeneous. Here, the authors create chamber-specific human pluripotent stem cell-derived cardiomyocytes to form both ventricular and atrial cells before embedding in collagen-based matrix to form ring-shaped engineered heart tissue.
- Idit Goldfracht
- , Stephanie Protze
- & Lior Gepstein
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Restoration of visual function by transplantation of optogenetically engineered photoreceptors
A challenge in cell therapy for photoreceptor degeneration is inducing the grafted cells to grow and maintain light sensitive outer segments. Here, the authors use optogenetics to circumvent this problem and restore visual function in blind mice.
- Marcela Garita-Hernandez
- , Maruša Lampič
- & Jens Duebel
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YAP inhibition enhances the differentiation of functional stem cell-derived insulin-producing β cells
Pluripotent stem cells can be directed into insulin-producing beta cells in vitro. Here, the authors show that downregulation of YAP, an effector of the Hippo pathway, enhances endocrine progenitor differentiation and the generation of beta-cells with improved insulin secretion.
- Edwin A. Rosado-Olivieri
- , Kendall Anderson
- & Douglas A. Melton
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Precisely controlling endogenous protein dosage in hPSCs and derivatives to model FOXG1 syndrome
Altered dosage of developmental regulators such as transcription factors can result in disorders, such as FOXG1 syndrome. Here, the authors demonstrate the utility of SMASh technology for modulating protein dosage by modeling FOXG1 syndrome using human pluripotent stem cell-derived neurons and neural organoids.
- Wenliang Zhu
- , Boya Zhang
- & Baoyang Hu
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Article
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Pluripotency and X chromosome dynamics revealed in pig pre-gastrulating embryos by single cell analysis
Lineage segregation from conception to gastrulation has been mapped at the single cell level in mouse, human and monkey. Here, the authors provide a comprehensive analysis of porcine preimplantation development using single cell RNA-seq; mapping metabolic changes, X chromosome inactivation and signalling pathways.
- Priscila Ramos-Ibeas
- , Fei Sang
- & Ramiro Alberio
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Bioreactor-based mass production of human iPSC-derived macrophages enables immunotherapies against bacterial airway infections
Pulmonary infections constitute a substantial health problem worldwide. Here the authors show that phagocytes similar to primitive macrophages can be generated from human induced pluripotent stem cells, by the use of industry-compatible, stirred-tank bioreactors, and applied as a cell-based therapy to treat acute bacterial infections in mice.
- Mania Ackermann
- , Henning Kempf
- & Nico Lachmann
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BMI1 enables interspecies chimerism with human pluripotent stem cells
Conventional human pluripotent stem cells (hPSCs) fail to contribute to interspecies chimaeras when injected into mouse blastocysts. Here the authors show that forced expression of BMI1 overcomes apoptosis of hPSCs in blastocysts of mouse, rabbit and pig allowing them to contribute to chimaeras.
- Ke Huang
- , Yanling Zhu
- & Guangjin Pan
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Generation of orthotopically functional salivary gland from embryonic stem cells
Functional salivary glands have not been generated from embryonic stem cells (mESCs) to date. Here the authors demonstrate directed in vitro differentiation of mESCs to oral ectoderm and salivary gland rudiments that form mature, functional salivary glands after orthotopic transplantation.
- Junichi Tanaka
- , Miho Ogawa
- & Kenji Mishima
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Interleukin-2 induces the in vitro maturation of human pluripotent stem cell-derived intestinal organoids
Human pluripotent stem cell-derived intestinal organoids (hIOs) are a useful model with which to study intestinal development and disease, but they require in vivo maturation to resemble adult tissue. Here, the authors show that T lymphocyte-derived IL-2 induces hIO maturation in vitro through the activation of STAT3.
- Kwang Bo Jung
- , Hana Lee
- & Mi-Young Son
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Human pluripotent reprogramming with CRISPR activators
CRISPRa is an attractive tool for cellular reprogramming due to its multiplexing capacity and direct targeting of genomic loci. Here the authors demonstrate the reprogramming of human fibroblasts into iPSCs, which is enhanced by targeting a conserved Alu-motif.
- Jere Weltner
- , Diego Balboa
- & Timo Otonkoski
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Targeting repair pathways with small molecules increases precise genome editing in pluripotent stem cells
Small molecule inhibitors can influence the choice of repair pathways, enhancing nucleotide substitution and gene integration in CRISPR-mediated genome editing. Here the authors introduce CRISPY, a mix of small molecules that can enhance precise editing with Cpf1 and Cas9D10A in hiPSCs.
- Stephan Riesenberg
- & Tomislav Maricic
Cellular reprogramming as a tool to model human aging in a dish