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Correction of Hirschsprung-associated mutations in human induced pluripotent stem cells via clustered regularly interspaced short palindromic repeats/cas9, restores neural crest cell function
Lai FP, Lau ST, Wong JK, Gui H, Wang RX, Zhou T, Lai WH, Tse HF, Tam PK, Garcia-Barcelo MM, Ngan ES (2017) Correction of Hirschsprung-associated mutations in human induced pluripotent stem cells via clustered regularly interspaced short palindromic repeats/cas9, restores neural crest cell function. Gastroenterology 153(1):139-153.e8. doi: 10.1053/j.gastro.2017.03.014 PMID: 28342760
Objective: To identify mutations associated with short-segment Hirschsprung disease (S-HSCR), and use the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing system to determine how mutations affect enteric neural crest cells (ENCC) function.
Summary: Mutations in vinculin gene (VCL) associated with S-HSCR were identified. Correction of this mutation in iPSC using CRISPR/Cas9 editing, as well as the RET G731del mutation that causes Hirschsprung disease with total colonic aganglionosis, restored ENCC function. The study demonstrates how human iPSCs can be used to identify disease-associated mutations and determine how they affect cell functions and contribute to pathogenesis.
Usage: Immunofluorescence (1:500)
Related Products: NGFR (mu p75) Rabbit Polyclonal, affinity-purified (Cat. #AB-N01AP)
Sox10+ adult stem cells contribute to biomaterial encapsulation and microvascularization
Wang D, Wang A, Wu F, Qiu X, Li Y, Chu J, Huang WC, Xu K, Gong X, Li S (2017) Sox10+ adult stem cells contribute to biomaterial encapsulation and microvascularization. Sci Rep 7:40295. doi: 10.1038/srep40295 PMID: 28071739
Objective: To show that Sox10+ adult stem cells contribute to both encapsulation and microvessel formation.
Summary: This study provides a novel mechanism that Sox10+ adult stem cells in the stroma of subcutaneous loose connective tissues are a common precursor of fibroblasts/myofibroblasts and perivascular cells. These adult stem cells can first differentiate into fibroblast-like cells at early stages of biomaterials implantation, and then into myofibroblasts promoting encapsulation/fibrosis, or perivascular cells supporting microvessels.
Usage: flow cytometry
Related Products: NGFR (mu p75) Rabbit Polyclonal, affinity-purified (Cat. #AB-N01AP)
Murine neural crest stem cells and embryonic stem cell-derived neuron precursors survive and differentiate after transplantation in a model of dorsal root avulsion
Konig N, Trolle C, Kapuralin K, Adameyko I, Mitrecic D, Aldskogius H, Shortland PJ, Kozlova EN (2017) Murine neural crest stem cells and embryonic stem cell-derived neuron precursors survive and differentiate after transplantation in a model of dorsal root avulsion. J Tissue Eng Regen Med 11(1):129-137. doi: 10.1002/term.1893 PMID: 24753366
Objective: To compare survival and migration of murine boundary cap neural crest stem cells (bNCSCs) and embryonic stem cells (ESCs)-derived, pre-differentiated neuron precursors after their implantation acutely at the junction between avulsed dorsal roots L3-L6 and the spinal cord.
Summary: The data show that both stem cell types successfully survived implantation to the acutely injured spinal cord and maintained their differentiation and migration potential. The data suggest that, depending on the source of neural stem cells, they can play different beneficial roles for recovery after dorsal root avulsion.
Usage: immunohistochemistry (1:500)
Related Products: NGFR (mu p75) Rabbit Polyclonal, affinity-purified (Cat. #AB-N01AP)
Placenta-derived mesenchymal stem cells facilitate neural and cognitive recovery in dementia rat model.
Cho J, Lee J, Jeong D, Kim H, Chang W, Moon J, Chang J (2016) Placenta-derived mesenchymal stem cells facilitate neural and cognitive recovery in dementia rat model. Neuroscience 2016 Abstracts 38.10 / G29. Society for Neuroscience, San Diego, CA.
Summary: Introduction: Dementia is a term that encompasses various types of neurodegenerative diseases of the brain that cause a gradual decline in mental abilities. Loss of cholinergic neurons in the brain cholinergic system including the hippocampus is a hallmark of many dementia cases. In this study, we report the therapeutic effects of administration of human placenta-derived mesenchymal stem cells (pMSCs) in dementia model Sprague-Dawley (SD) rats using two different cell injection methods: intracerebroventricular (ICV) and intravenous (IV) injections. Methods: Dementia modeling was carried out by intraventricular injection of 192 IgG saporin, which causes lesion of cholinergic neurons. Fifty male SD rats were divided into four groups: normal (n=9), lesion (n=9), ICV (n=12) and IV (n=12). All rats were then subject to Morris water maze test and subsequent immunostaining analyses using markers for human cytoplasm, acetylcholinesterase (AChE), choline acetyltransferase (ChAT) and microglial cells at the hippocampus. Results: Lesioned rats showed poor performance in the Morris water maze test compared to the normal rats. Both ICV and IV pMSC administration allowed significant cognitive recovery compared to the lesioned rats. AChE was also significantly recovered back to normal levels at the hippocampus in rats injected with pMSCs post-lesion. ChAT did not co-localize with pMSCs, showing that pMSCs did not directly differentiate into cholinergic cells. Stem cell count showed a significantly greater number of pMSCs at the hippocampal dentate gyrus in IV group rats compared to ICV group rats. Number of microglial cells increased in lesioned rats, and was significantly reduced back to normal levels after pMSC injection. Discussion: Our results demonstrate that injection of pMSCs facilitates recovery of cholinergic neuronal population and function, as well as cognitive behavior. The mechanism through which such recovery happens does not seem to be direct differentiation of injected pMSCs into cholinergic neurons, but rather seems to be through paracrine effects that resemble microglial function. Further research will be necessary for elucidation of the exact mechanisms involved and establishment of optimal parameters for successful cell homing. Acknowledgements: This study was supported by the grant from the Yonsei University Future-leading Research Initiative (Yonsei Challenge) of 2015 (2015-22-0137) and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (2015R1C1A1A02036851).
Related Products: 192-IgG-SAP (Cat. #IT-01)
Derivation of corneal keratocyte-like cells from human induced pluripotent stem cells.
Naylor R, McGhee C, Cowan C, Davidson A, Holm T, Sherwin T (2016) Derivation of corneal keratocyte-like cells from human induced pluripotent stem cells. PLoS One 11:e0165464. doi: 10.1371/journal.pone.0165464 PMID: 27792791
Summary: Slides containing cryosections were dried overnight at 4°C and then washed twice in Tris Buffered Saline containing 0.1% Triton X100 (TBST). Slides were then placed in block solution (3% BSA, 5% Goat serum in TBST) for at least one hour. The primary antibody was then applied in the same block solution (1:100) and left overnight at 4°C.
Related Products: NGFr (ME20.4, p75) Mouse Monoclonal (Cat. #AB-N07)
Stem cells in tissue regeneration and diseases
Huang WC (2016) Stem cells in tissue regeneration and diseases. University of California, Berkeley Thesis.
Objective: To investigate the therapeutic effect of induced pluripotent stem cell-derived neural crest stem cells (iPSC-NCSCs) and mesenchymal progenitor cells (MPCs) on peripheral nerve regeneration.
Summary: Transplantation of NCSCs has better outcomes of motor nerve recovery and muscle reinnervation by Schwann cell differentiation in vivo and paracrine signaling, whereas transplantation of MPCs fails to promote functional nerve regeneration.
Usage: Immunostaining to characterize MPCs
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Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin.
Palchaudhuri R, Saez B, Hoggatt J, Schajnovitz A, Sykes D, Tate T, Czechowicz A, Kfoury Y, Ruchika F, Rossi D, Verdine G, Mansour M, Scadden D (2016) Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol 34:738-745. doi: 10.1038/nbt.3584
Summary: To demonstrate correction of a clinically relevant disease, we employed CD45-SAP in a mouse model of sickle cell anemia and demonstrated our method achieved >90% donor cell chimerism, all mice in three groups (18/18), resulting in complete disease correction (red blood cell counts, hemoglobin levels, hematocrit levels and reticulocyte frequencies were returned to normal). If these pre-clinical results can be successfully translated to the clinic, it would greatly reduce conditioning-related toxicities and expand the use of hematopoietic stem cell transplantation.
Related Products: Streptavidin-ZAP (Cat. #IT-27), Anti-CD45.2-SAP (Cat. #IT-91)
Featured Article: Targeted depletion of hematopoietic stem cells promises safer transplantation
Palchaudhuri R (2016) Featured Article: Targeted depletion of hematopoietic stem cells promises safer transplantation. Targeting Trends 17(3)
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Read the featured article in Targeting Trends.
See Also:
- Palchaudhuri R et al. Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol 34:738-745, 2016.
- Skulimowska I et al. The biology of hematopoietic stem cells and its clinical implications. FEBS J 16192, 2021.
Differentiation defect in neural crest-derived smooth muscle cells in patients with aortopathy associated with bicuspid aortic valves.
Jiao J, Xiong W, Wang L, Yang J, Qiu P, Hirai H, Shao L, Milewicz D, Chen Y, Yang B (2016) Differentiation defect in neural crest-derived smooth muscle cells in patients with aortopathy associated with bicuspid aortic valves. EBioMedicine 10:282-290. doi: 10.1016/j.ebiom.2016.06.045 PMID: 27394642
Summary: Individuals with bicuspid aortic valves (BAV) are at a higher risk of developing thoracic aortic aneurysms (TAA) than patients with trileaflet aortic valves (TAV). Aneurysms associated with BAV most commonly involve the ascending aorta. Smooth muscle cells (SMCs) in the ascending and descending aorta arise from neural crest (NC) and paraxial mesoderm (PM), respectively. Scientists hypothesized defective differentiation of the neural crest stem cells (NCSCs)-derived SMCs but not paraxial mesoderm cells (PMCs)- derived SMCs contributes to the aortopathy associated with BAV. Induced pluripotent stem cells (iPSCs) from BAV/TAA patients were differentiated into NCSC-derived SMCs and showed decreased expression of a marker of SMC differentiation (MYH11) and impaired contraction. The scientists demonstrated that decreased differentiation and contraction of patient’s NCSC-derived SMCs may contribute to the aortopathy associated with BAV.
Usage: Anti-NGFr (ME20.4, p75, Cat. #AB-N07) was used for the immunofluorescence staining and flow cytometry of NCSCs.
Related Products: NGFr (ME20.4, p75) Mouse Monoclonal (Cat. #AB-N07)
Bone morphogenetic protein 4 promotes craniofacial neural crest induction from human pluripotent stem cells.
Mimura S, Suga M, Okada K, Kinehara M, Nikawa H, Furue M (2016) Bone morphogenetic protein 4 promotes craniofacial neural crest induction from human pluripotent stem cells. Int J Dev Biol 60:21-28. doi: 10.1387/ijdb.160040mk PMID: 26934293
Usage: Immunocytochemistry and flow cytometry
Related Products: NGFr (ME20.4, p75) Mouse Monoclonal (Cat. #AB-N07)