scholarly journals An Efficient Protocol for Single-Cell Cloning Human Pluripotent Stem Cells

2019 ◽  
Vol 7 ◽  
Author(s):  
Amar M. Singh
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Cell Cloning ◽  
Single Cell Cloning

Single-cell cloning and expansion of human induced pluripotent stem cells by a microfluidic culture device

2014 ◽  
Vol 453 (1) ◽  
pp. 131-137 ◽  
Author(s):  
Taku Matsumura ◽  
Kazuya Tatsumi ◽  
Yuichiro Noda ◽  
Naoyuki Nakanishi ◽  
Atsuhito Okonogi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Cell Cloning ◽  
Single Cell Cloning ◽  
Induced Pluripotent

scholarly journals Development of an efficient single-cell cloning and expansion strategy for genome edited induced pluripotent stem cells

2021 ◽  
Author(s):  
Nupur Bhargava ◽  
Priya Thakur ◽  
Thulasi Priyadharshini Muruganandam ◽  
Shashank Jaitly ◽  
Pragya Gupta ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Viability ◽  
Single Cell ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Genetic Mutations ◽  
Disease Models ◽  
Cell Cloning ◽  
Single Cell Cloning ◽  
Induced Pluripotent

Disease-specific human induced pluripotent stem cells (hiPSCs) can be generated directly from individuals with known disease characteristics or alternatively be modified using genome editing approaches to introduce disease causing genetic mutations to study the biological response of those mutations. The genome editing procedure in hiPSCs is still inefficient, particularly when it comes to homology directed repair (HDR) of genetic mutations or targeted transgene insertion in the genome and single cell cloning of edited cells. In addition, genome editing processes also involve additional cellular stresses such as trouble with cell viability and genetic stability of hiPSCs. Therefore, efficient workflows are desired to increase genome editing application to hiPSC disease models and therapeutic applications. Apart from genome editing efficiency, hiPSC survival following single-cell cloning has proved to be challenging and has thus restricted the capability to easily isolate homogeneous clones from edited hiPSCs. To this end, we demonstrate an efficient workflow for feeder-free single cell clone generation and expansion in both CRISPR-mediated knock-out (KO) and knock-in (KI) hiPSC lines. Using StemFlex medium and CloneR supplement in conjunction with Matrigel cell culture matrix, we show that cell viability and expansion during single-cell cloning in edited and unedited cells is significantly enhanced. Our reliable single-cell cloning and expansion workflow did not affect the biology of the hiPSCs as the cells retained their growth and morphology, expression of various pluripotency markers and normal karyotype. This simplified and efficient workflow will allow for a new level of sophistication in generating hiPSC-based disease models to promote rapid advancement in basic research and also the development of novel cellular therapeutics.

scholarly journals Single-Cell Analysis Reveals Partial Reactivation of X Chromosome instead of Chromosome-wide Dampening in Naive Human Pluripotent Stem Cells

2020 ◽  
Vol 14 (5) ◽  
pp. 745-754 ◽  
Author(s):  
Susmita Mandal ◽  
Deepshikha Chandel ◽  
Harman Kaur ◽  
Sudeshna Majumdar ◽  
Maniteja Arava ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
X Chromosome ◽  
Pluripotent Stem Cells ◽  
Single Cell Analysis ◽  
Human Pluripotent Stem Cells ◽  
Cell Analysis

scholarly journals Transcriptional Heterogeneity in Naive and Primed Human Pluripotent Stem Cells at Single-Cell Resolution

Cell Reports ◽  
2019 ◽  
Vol 26 (4) ◽  
pp. 815-824.e4 ◽  
Author(s):  
Tobias Messmer ◽  
Ferdinand von Meyenn ◽  
Aurora Savino ◽  
Fátima Santos ◽  
Hisham Mohammed ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells

scholarly journals Methods for Automated Single Cell Isolation and Sub‐Cloning of Human Pluripotent Stem Cells

2020 ◽  
Vol 55 (1) ◽  
Author(s):  
Valeria Fernandez Vallone ◽  
Narasimha Swamy Telugu ◽  
Iris Fischer ◽  
Duncan Miller ◽  
Sandra Schommer ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Cell Isolation ◽  
Human Pluripotent Stem Cells ◽  
Single Cell Isolation

scholarly journals Single-Cell Gene Expression Profiles Define Self-Renewing, Pluripotent, and Lineage Primed States of Human Pluripotent Stem Cells

2014 ◽  
Vol 2 (6) ◽  
pp. 881-895 ◽  
Author(s):  
Shelley R. Hough ◽  
Matthew Thornton ◽  
Elizabeth Mason ◽  
Jessica C. Mar ◽  
Christine A. Wells ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Human Pluripotent Stem Cells ◽  
Cell Gene Expression ◽  
Cell Gene

scholarly journals Single cell heterogeneity in human pluripotent stem cells

BMB Reports ◽  
2021 ◽  
Vol 54 (10) ◽  
pp. 505-515
Author(s):  
Seungbok Yang ◽  
Yoonjae Cho ◽  
Jiwon Jang
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Cell Heterogeneity

scholarly journals Single-Cell-State Culture of Human Pluripotent Stem Cells Increases Transfection Efficiency

2016 ◽  
Vol 5 (1) ◽  
pp. 127-136 ◽  
Author(s):  
Takenobu Nii ◽  
Hiroshi Kohara ◽  
Tomotoshi Marumoto ◽  
Tetsushi Sakuma ◽  
Takashi Yamamoto ◽  
...  
Keyword(s):  
Stem Cells ◽  
Single Cell ◽  
Pluripotent Stem Cells ◽  
Transfection Efficiency ◽  
Human Pluripotent Stem Cells ◽  
State Culture ◽  
Cell State
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