Publications by authors named "Suchan Niroula"

3 Publications

  • Page 1 of 1

Protocol for Cloning Epithelial Stem Cell Variants from Human Lung.

STAR Protoc 2020 Sep 9;1(2). Epub 2020 Jul 9.

Stem Cell Center, Department of Biology and Biochemistry, University of Houston, Houston, TX 77204, USA.

The plurality of clonogenic cells derived from human lung includes a spectrum of diverse p63+ stem cells responsible for the regeneration of normal epithelial tissue and disease-associated metaplastic lesions. Here, we report protocols for the cloning, expansion, and characterization of these stem cell variants, which in general assist in analyses of stem cell heterogeneity, genome editing, drug screening, and regenerative medicine. For complete details on the use and execution of this protocol, please refer to Kumar et al. (2011), Zuo et al. (2015), and Rao et al. (2020).
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http://dx.doi.org/10.1016/j.xpro.2020.100063DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7529324PMC
September 2020

Regenerative Metaplastic Clones in COPD Lung Drive Inflammation and Fibrosis.

Cell 2020 05 15;181(4):848-864.e18. Epub 2020 Apr 15.

Stem Cell Center, Department of Biology and Biochemistry, University of Houston, Houston, TX 77003, USA. Electronic address:

Chronic obstructive pulmonary disease (COPD) is a progressive condition of chronic bronchitis, small airway obstruction, and emphysema that represents a leading cause of death worldwide. While inflammation, fibrosis, mucus hypersecretion, and metaplastic epithelial lesions are hallmarks of this disease, their origins and dependent relationships remain unclear. Here we apply single-cell cloning technologies to lung tissue of patients with and without COPD. Unlike control lungs, which were dominated by normal distal airway progenitor cells, COPD lungs were inundated by three variant progenitors epigenetically committed to distinct metaplastic lesions. When transplanted to immunodeficient mice, these variant clones induced pathology akin to the mucous and squamous metaplasia, neutrophilic inflammation, and fibrosis seen in COPD. Remarkably, similar variants pre-exist as minor constituents of control and fetal lung and conceivably act in normal processes of immune surveillance. However, these same variants likely catalyze the pathologic and progressive features of COPD when expanded to high numbers.
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http://dx.doi.org/10.1016/j.cell.2020.03.047DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7294989PMC
May 2020

Cloning of ground-state intestinal stem cells from endoscopic biopsy samples.

Nat Protoc 2020 05 1;15(5):1612-1627. Epub 2020 Apr 1.

Stem Cell Center, Department of Biology and Biochemistry, University of Houston, Houston, Texas, USA.

'Adult' or 'somatic' stem cells harbor an intrinsic ability to regenerate tissues. Heterogeneity of such stem cells along the gastrointestinal tract yields the known segmental specificity of this organ and may contribute to the pathology of certain enteric conditions. Here we detail technology for the generation of 'libraries' of clonogenic cells from 1-mm-diamter endoscopic biopsy samples from the human gastrointestinal tract. Each of the 150-300 independent clones in a typical stem cell library can be clonally expanded to billions of cells in a few weeks while maintaining genomic stability and the ability to undergo multipotent differentiation to the specific epithelia from which the sample originated. The key to this methodology is the intrinsic immortality of normal intestinal stem cells (ISCs) and culture systems that maintain them as highly immature, ground-state ISCs marked by a single-cell clonogenicity of 70% and a corresponding 250-fold proliferative advantage over spheroid technologies. Clonal approaches such as this enhance the resolution of molecular genetics, make genome editing easier, and may be useful in regenerative medicine, unravelling heterogeneity in disease, and facilitating drug discovery.
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http://dx.doi.org/10.1038/s41596-020-0298-4DOI Listing
May 2020