scholarly journals Induction of Sertoli cells from human fibroblasts by NR5A1 and GATA4

2018 ◽  
Author(s):  
Jianlin Liang ◽  
Nan Wang ◽  
Jing He ◽  
Jian Du ◽  
Yahui Guo ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Lipid Droplets ◽  
Human Fibroblasts ◽  
Seminiferous Tubules ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
Reprogramming Factors ◽  
Spermatogonia Cells

AbstractSertoli cells are essential nurse cells in the testis that regulate the process of spermatogenesis and establish the immune-privileged environment of the blood-testis-barrier (BTB). The induction of human Sertoli cells from fibroblasts could provide cellular sources for fertility and transplantation treatments. Here, we report the in vitro reprogramming of human fibroblasts to Sertoli cells and characterize these human induced Sertoli cells (hiSCs). Initially, five transcriptional factors (NR5A1, GATA4, WT1, SOX9 and DMRT1) and a gene reporter carrying the AMH promoter were utilized to obtain the hiSCs. We further reduce the number of reprogramming factors to two, i.e., NR5A1 and GATA4, and show that these hiSCs have transcriptome profiles that are similar to those of primary human Sertoli cells. Consistent with the known cellular properties of Sertoli cells, hiSCs attract endothelial cells and exhibit high number of lipid droplets in the cytoplasm. More importantly, hiSCs can sustain the viability of spermatogonia cells harvested from mouse seminiferous tubules. In addition, hiSCs suppress the production of IL-2 and proliferation of human T lymphocytes. When hiSCs were cotransplanted with human embryonic kidney cells, these xenotransplanted human cells survived longer in mice with normal immune systems. hiSCs also allow us to determine a gene associated with Sertoli-only syndrome (SCO), CX43, is indeed important in regulating the maturation of Sertoli cells.

scholarly journals Induction of Sertoli-like cells from human fibroblasts by NR5A1 and GATA4

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Jianlin Liang ◽  
Nan Wang ◽  
Jing He ◽  
Jian Du ◽  
Yahui Guo ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Sertoli Cell ◽  
Sertoli Cells ◽  
Human Fibroblasts ◽  
Seminiferous Tubules ◽  
Nurse Cells ◽  
Immune Systems ◽  
Reprogramming Factors ◽  
Spermatogonia Cells

Sertoli cells are essential nurse cells in the testis that regulate the process of spermatogenesis and establish the immune-privileged environment of the blood-testis-barrier (BTB). Here, we report the in vitro reprogramming of fibroblasts to human induced Sertoli-like cells (hiSCs). Initially, five transcriptional factors and a gene reporter carrying the AMH promoter were utilized to obtain the hiSCs. We further reduce the number of reprogramming factors to two, NR5A1 and GATA4, and show that these hiSCs have transcriptome profiles and cellular properties that are similar to those of primary human Sertoli cells. Moreover, hiSCs can sustain the viability of spermatogonia cells harvested from mouse seminiferous tubules. hiSCs suppress the proliferation of human T lymphocytes and protect xenotransplanted human cells in mice with normal immune systems. hiSCs also allow us to determine a gene associated with Sertoli cell only syndrome (SCO), CX43, is indeed important in regulating the maturation of Sertoli cells.

Molecular insight to in vitro biocompatibility of phytofabricated copper oxide nanoparticles with human embryonic kidney cells

Nanomedicine ◽  
2018 ◽  
Vol 13 (19) ◽  
pp. 2415-2433 ◽  
Author(s):  
Puja Kumari ◽  
Pritam Kumar Panda ◽  
Ealisha Jha ◽  
Nandini Pramanik ◽  
Kumari Nisha ◽  
...  
Keyword(s):  
Copper Oxide ◽  
Copper Oxide Nanoparticles ◽  
Oxide Nanoparticles ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
In Vitro Biocompatibility

scholarly journals A novel UBA and UBX domain protein that binds polyubiquitin and VCP and is a substrate for SAPKs

2004 ◽  
Vol 384 (2) ◽  
pp. 391-400 ◽  
Author(s):  
Helen McNEILL ◽  
Axel KNEBEL ◽  
J. Simon C. ARTHUR ◽  
Ana CUENDA ◽  
Philip COHEN
Keyword(s):  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Glutathione S Transferase ◽  
Hek 293 ◽  
Human Embryonic Kidney Cells ◽  
Extracellular Signal Regulated Kinase ◽  
Cell Extracts ◽  
Extracellular Signal ◽  
Sb 203580

A widely expressed protein containing UBA (ubiquitin-associated) and UBX (ubiquitin-like) domains was identified as a substrate of SAPKs (stress-activated protein kinases). Termed SAKS1 (SAPK substrate-1), it was phosphorylated efficiently at Ser200in vitro by SAPK3/p38γ, SAPK4/p38δ and JNK (c-Jun N-terminal kinase), but weakly by SAPK2a/p38α, SAPK2b/p38β2 or ERK (extracellular-signal-regulated kinase) 2. Ser200, situated immediately N-terminal to the UBX domain, became phosphorylated in HEK-293 (human embryonic kidney) cells in response to stressors. Phosphorylation was not prevented by SB 203580 (an inhibitor of SAPK2a/p38α and SAPK2b/p38β2) and/or PD 184352 (which inhibits the activation of ERK1 and ERK2), and was similar in fibroblasts lacking both SAPK3/p38γ and SAPK4/p38δ or JNK1 and JNK2. SAKS1 bound ubiquitin tetramers and VCP (valosin-containing protein) in vitro via the UBA and UBX domains respectively. The amount of VCP in cell extracts that bound to immobilized GST (glutathione S-transferase)–SAKS1 was enhanced by elevating the level of polyubiquitinated proteins, while SAKS1 and VCP in extracts were coimmunoprecipitated with an antibody raised against S5a, a component of the 19 S proteasomal subunit that binds polyubiquitinated proteins. PNGase (peptide N-glycanase) formed a 1:1 complex with VCP and, for this reason, also bound to immobilized GST–SAKS1. We suggest that SAKS1 may be an adaptor that directs VCP to polyubiquitinated proteins, and PNGase to misfolded glycoproteins, facilitating their destruction by the proteasome.

scholarly journals The effect of simvastatin on lipid droplets accumulation in human embryonic kidney cells and pancreatic cancer cells

2013 ◽  
Vol 12 (1) ◽  
Author(s):  
Helena Gbelcová ◽  
Martin Švéda ◽  
Lucia Laubertová ◽  
Ivan Varga ◽  
Libor Vítek ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Cells ◽  
Lipid Droplets ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
Pancreatic Cancer Cells

Generation of Human Embryonic Kidney Cells with Extended In Vitro Life Span through Viral Oncogene Transfection

1989 ◽  
Vol 7 (9) ◽  
pp. 939-946 ◽  
Author(s):  
Steve Abcouwer ◽  
Polly S. Robinson ◽  
Charles F. Goochee ◽  
Michael T. Crow
Keyword(s):  
Life Span ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
Viral Oncogene

Hyperthermia with Magnetic Nanowires for Inactivating Living Cells

2008 ◽  
Vol 8 (5) ◽  
pp. 2323-2327 ◽  
Author(s):  
D. S. Choi ◽  
J. Park ◽  
S. Kim ◽  
D. H. Gracias ◽  
M. K. Cho ◽  
...  
Keyword(s):  
Cell Death ◽  
Magnetic Moments ◽  
Magnetic Hysteresis ◽  
Kidney Cells ◽  
Magnetic Nanowires ◽  
Human Embryonic Kidney ◽  
Lower Field ◽  
Hek 293 ◽  
Human Embryonic Kidney Cells

We describe a method to induce hyperthermia in cells, in-vitro, by remotely heating Ni nanowires (NWs) with radio frequency (RF) electromagnetic fields. Ni NWs were internalized by human embryonic kidney cells (HEK-293). Only cells proximal to NWs or with internalized NWs changed shape on exposure to RF fields indicative of cell death. The cell death occurs as a result of hyperthermia, since the RF field remotely heats the NWs as a result of magnetic hysteresis. This is the first demonstration of hyperthermia induced by NWs; since the NWs have anisotropic and strong magnetic moments, our experiments suggest the possibility of performing hyperthermia at lower field strengths in order to minimize damage to untargeted cells in applications such as the treatment of cancer.

In vitro renal toxicity evaluation of copper-based metal–organic framework HKUST-1 on human embryonic kidney cells

2021 ◽  
Vol 273 ◽  
pp. 116528
Author(s):  
Yi-Chun Chen ◽  
Kun-Yi Andrew Lin ◽  
Ku-Fan Chen ◽  
Xin-Yu Jiang ◽  
Chia-Hua Lin
Keyword(s):  
Renal Toxicity ◽  
Metal Organic Framework ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Toxicity Evaluation ◽  
Human Embryonic Kidney Cells ◽  
Metal Organic ◽  
Organic Framework

scholarly journals In vitro transcribed guide RNAs trigger an innate immune response via the RIG-I pathway

2018 ◽  
Author(s):  
Beeke Wienert ◽  
Jiyung Shin ◽  
Elena Zelin ◽  
Kathleen Pestal ◽  
Jacob E. Corn
Keyword(s):  
Immune Response ◽  
Genome Editing ◽  
Innate Immune ◽  
Kidney Cells ◽  
Primary Cells ◽  
Human Embryonic Kidney ◽  
Hematopoietic Stem ◽  
Human Embryonic Kidney Cells ◽  
Guide Rna

AbstractCRISPR-Cas9 genome editing is revolutionizing fundamental research and has great potential for the treatment of many diseases. While editing of immortalized cell lines has become relatively easy, editing of therapeutically relevant primary cells and tissues can remain challenging. One recent advancement is the delivery of a Cas9 protein and an in vitro transcribed (IVT) guide RNA (gRNA) as a precomplexed ribonucleoprotein (RNP). This approach allows editing of primary cells such as T cells and hematopoietic stem cells, but the consequences beyond genome editing of introducing foreign Cas9 RNPs into mammalian cells are not fully understood. Here we show that the IVT gRNAs commonly used by many laboratories for RNP editing trigger a potent innate immune response that can be several thousand times stronger than benchmark immune stimulating ligands. IVT gRNAs are recognized in the cytosol through the RIG-I pathway but not the MDA5 pathway, thereby triggering a type I interferon response. Removal of the 5’-triphosphate from gRNAs ameliorates inflammatory signaling and prevents the loss of viability associated with genome editing in hematopoietic stem cells. The potential for Cas9 RNP editing to induce a potent antiviral response indicates that care must be taken when designing therapeutic strategies to edit primary cells.AbbreviationsCasCRISPR-associatedCIPcalf intestinal alkaline phosphataseCRISPRclustered, regularly interspaced, short palindromic repeatdCas9nuclease-dead Cas9HEK293Human embryonic kidney cells 293HEK293THuman embryonic kidney cells 293 SV40 large T antigenHeLaHenrietta Lacks cellsHSPCsCD34+ human hematopoietic stem and progenitor cellsIFNAR1Interferon Alpha And Beta Receptor Subunit 1IFNβ/IFNB1Interferon betaISG15Interferon-stimulated gene 15 IVT – in vitro transcribedKOknockoutMAVSmitochondrial activator of virus signalingMDA5/IFIH1melanoma differentiation-associated gene 5/ Interferon Induced with Helicase C Domain 1PAMPpathogen-associated molecular patternRIG-I/DDX58retinoic acid-inducible gene I/ DExD-H-box helicase 58gRNAguide RNASPRIsolid phase reversible immobilizationWTwild type

scholarly journals Cytotoxic Aporphines from Artabotrys Crassifolius

2016 ◽  
Vol 11 (3) ◽  
pp. 1934578X1601100
Author(s):  
Tan Kok Kwan ◽  
Fiona Shipton ◽  
Nadiah Syafiqah Nor Azman ◽  
Shahadat Hossan ◽  
Khoo Ten Jin ◽  
...  
Keyword(s):  
Chloroform Extract ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Cytotoxic Effects ◽  
Human Embryonic Kidney Cells ◽  
Hct 116 ◽  
Ethanol Extracts ◽  
Hct 116 Cells ◽  
Mcf 7

Artabotrys crassifolius Hook. f. & Thomson is a medicinal plant used in Malaysia. The cytotoxic effects of the hexane, chloroform and ethanol extracts of the leaves and bark were examined in vitro against MCF-7, MDA-468 and HCT-116 cells. The chloroform extract of the bark inhibited the growth of all cell lines with GI50 values ranging from 4.2 μg/mL to 9.4 μg/mL. Silica gel column chromatography of this extract yielded artabotrine, liridine, atherospermidine and lysicamine. Artabotrine and lysicamine inhibited the growth of HCT-116 and MCF-7 cells with GI50 values ranging from 3.3 μM to 3.9 μM. These alkaloids were not toxic to human embryonic kidney cells (HEK297) up to a concentration of 50 μg/mL.

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