Using Laser Direct-Write to Influence Embryoid Body Size on Uniform Substrates

2012 ◽  
Author(s):  
Andrew D. Dias ◽  
Yubing Xie ◽  
Douglas B. Chrisey ◽  
David T. Corr
Keyword(s):  
Stem Cells ◽  
Body Size ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Direct Write ◽  
Cell Type ◽  
Differentiation Potential ◽  
Self Renewal ◽  
Cell Sources ◽  
Laser Direct Write

Embryonic stem cells (ESCs) are capable of self-renewal and have the potential to differentiate into any specialized cell type. Understanding and directing this differentiation potential is critical to producing cell sources for replacement therapies.

scholarly journals The microwell control of embryoid body size in order to regulate cardiac differentiation of human embryonic stem cells

Biomaterials ◽  
2010 ◽  
Vol 31 (7) ◽  
pp. 1885-1893 ◽  
Author(s):  
Jeffrey C. Mohr ◽  
Jianhua Zhang ◽  
Samira M. Azarin ◽  
Andrew G. Soerens ◽  
Juan J. de Pablo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Body Size ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Cardiac Differentiation

Laser Direct-Write of Embryonic Stem Cells and Cells Encapsulated in Alginate Beads for Engineered Biological Constructs

2012 ◽  
Vol 1418 ◽  
Author(s):  
T.B. Phamduy ◽  
A.D. Dias ◽  
N. Abdul Raof ◽  
N.R. Schiele ◽  
D.T. Corr ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Alginate Beads ◽  
Stem Cell Marker ◽  
Direct Write ◽  
Pluripotency Marker ◽  
Single Bead ◽  
Substrate Surfaces ◽  
Laser Direct Write

AbstractThe ability to control the deposition of mouse embryonic stem cells (mESCs), and mESCs encapsulated in 200-μm diameter alginate microbeads, into customized patterns has recently been achieved using laser direct-write (LDW). Gelatin-based LDW was utilized to target and reproducibly deposit groups of cells directly onto receiving substrate surfaces. Live/dead staining for cell viability and immunocytochemistry for the pluripotency marker, Oct-4, indicated that transferred mESCs were viable following transfer, and maintained an important embryonic stem cell marker, respectively. LDW was further used to print mESCs encapsulated in hydrogel microbeads into customized patterns on a single-bead basis. Recent efforts have also achieved patterns of discrete co-cultures of mESCs and breast cancer cells in separate hydrogel microbeads. Altogether, we demonstrated the feasibility of LDW to print patterns of mESCs and mESC-microbeads for the biomimetic assembly of engineered cellular constructs and tissue models.

Precise Patterning of Mouse Embryonic Stem Cells on Glass Cover Slips for High-Throughput Analysis Using Laser Direct-Write

2011 ◽  
Author(s):  
Andrew D. Dias ◽  
Nathan R. Schiele ◽  
Brendan M. Carr ◽  
Nurazhani Abdul Raof ◽  
Yubing Xie ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
Embryonic Stem ◽  
Direct Write ◽  
Glass Cover ◽  
Computer Aided ◽  
Petri Dishes ◽  
Laser Direct Write

Engineering a microenvironment where the growth substrate and distance between cells are controlled is highly desirable to understand how cellular interactions affect stem cell differentiation. Laser direct-write (LDW) allows rapid and precise placement of living cells via computer-aided design/computer-aided manufacturing (CAD/CAM) control. Application of this technique to study the effects of various stem cell microenvironments on differentiation requires a high-throughput experimental setup [1]. Recently, our lab has developed a gelatin-based LDW method for the precise patterning of sensitive cell types, such as mouse embryonic stem cells (mESCs), at a resolution of about 5 μm [2]. Although viable mESCs were successfully printed with maintained pluripotency, this technique required cells to be patterned onto polystyrene Petri dishes [2,3], which may limit high-throughput efficiency. Moreover, the use of polystyrene Petri dishes requires large quantities of culture medium and is not convenient for biological analysis of mESC differentiation. Therefore, the objective of this study was to adapt the LDW method, without altering its prior success, to transfer patterns of viable mESCs to glass cover slips. However, this adaptation to cover slips could not be achieved through simple downscaling due to the unique challenges of providing sufficient moisture for viable cell transfer while maintaining pattern registry on a cover slip. Once cells have been laser patterned, cover slips can then be moved to a 24-well plate so that separate sets of laser patterned cells can be analyzed in parallel for higher experimental throughput utilizing fewer resources to maintain the cells.

Expression levels and activation status of Yap splicing isoforms determine self‐renewal and differentiation potential of embryonic stem cells

Stem Cells ◽  
2021 ◽  
Vol 39 (9) ◽  
pp. 1178-1191
Author(s):  
Xueyue Wang ◽  
Yan Ruan ◽  
Junlei Zhang ◽  
Yanping Tian ◽  
Lianlian Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Differentiation Potential ◽  
Self Renewal ◽  
Expression Levels ◽  
Splicing Isoforms ◽  
Activation Status

scholarly journals Hypoxic Culture Maintains Self-Renewal and Enhances Embryoid Body Formation of Human Embryonic Stem Cells

2010 ◽  
Vol 16 (9) ◽  
pp. 2901-2913 ◽  
Author(s):  
Hsin-Fu Chen ◽  
Hung-Chih Kuo ◽  
Shau-Ping Lin ◽  
Chung-Liang Chien ◽  
Ming-Shan Chiang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Self Renewal ◽  
Body Formation ◽  
Embryoid Body Formation

scholarly journals PS1 - 158 The Role of LIN28A in Neoplastic Transformation of Human Embryonic Stem Cells (hESCs)

2016 ◽  
Vol 43 (S4) ◽  
pp. S10-S10
Author(s):  
R. Kaur ◽  
L. Liang ◽  
T. Werbowetski-Ogilvie
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Rna Binding ◽  
Embryonic Stem ◽  
Neoplastic Transformation ◽  
Differentiation Potential ◽  
Self Renewal ◽  
Human Cancers

Human embryonic stem cells (hESCs) are known for their indefinite self-renewal ability and pluripotent nature. However, during long-term culture, normal hESCs can undergo neoplastic transformation and acquire enhanced self-renewal ability and aberrant differentiation potential. These transformed-hESCs (trans-hESCs) exhibit high expression of the pluripotent gene, LIN28A. LIN28A, an RNA binding protein, is known: for its role in self-renewal of hESCs, as a reprogramming factor for generating induced-pluripotent stem cells and as a potent oncogene in several poorly differentiated, highly malignant human cancers. Despite its multiple functions, how LIN28A contributes to neoplastic transformation of normal hESCs is poorly understood. Our preliminary data demonstrate that following LIN28A knockdown, trans-hESCs display normal hESCs morphology consisting of both pluripotent colony cells surrounded by more differentiated fibroblast-like cells. Neural precursors derived from LIN28A knockdown trans-hESCs also revert back to a state of normal cell morphology and growth. Further analyses revealed that the expression levels of stage-specific embryonic antigen (SSEA3), OCT3/4 and NANOG decreases and are comparable to that observed in normal hESCs following LIN28A downregulation. Expression of miRNA targets of LIN28A such as let7i and mir125b was increased to levels seen in normal hESCs. These preliminary results indicate that LIN28A is a major contributing factor to neoplastic transformation of hESCs and that this process can be reversed by cellular “reprogramming”. This study will enhance our understanding of role of LIN28A in the transformation process in various human cancers thus, underscoring the value of hESCs and their neoplastic-derivatives as cellular and molecular model for studying tumor progression.

scholarly journals The maintenance of pluripotency following laser direct-write of mouse embryonic stem cells

Biomaterials ◽  
2011 ◽  
Vol 32 (7) ◽  
pp. 1802-1808 ◽  
Author(s):  
Nurazhani Abdul Raof ◽  
Nathan R. Schiele ◽  
Yubing Xie ◽  
Douglas B. Chrisey ◽  
David T. Corr
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Direct Write ◽  
Laser Direct Write

scholarly journals Disruption of RING and PHD Domains of TRIM28 Evokes Differentiation in Human iPSCs

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1933
Author(s):  
Sylwia Mazurek ◽  
Urszula Oleksiewicz ◽  
Patrycja Czerwińska ◽  
Joanna Wróblewska ◽  
Marta Klimczak ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Self Renewal ◽  
Human Ipscs ◽  
New Gene ◽  
Induced Pluripotent ◽  
The Impact ◽  
Phd Domain

TRIM28, a multi-domain protein, is crucial in the development of mouse embryos and the maintenance of embryonic stem cells’ (ESC) self-renewal potential. As the epigenetic factor modulating chromatin structure, TRIM28 regulates the expression of numerous genes and is associated with progression and poor prognosis in many types of cancer. Because of many similarities between highly dedifferentiated cancer cells and normal pluripotent stem cells, we applied human induced pluripotent stem cells (hiPSC) as a model for stemness studies. For the first time in hiPSC, we analyzed the function of individual TRIM28 domains. Here we demonstrate the essential role of a really interesting new gene (RING) domain and plant homeodomain (PHD) in regulating pluripotency maintenance and self-renewal capacity of hiPSC. Our data indicate that mutation within the RING or PHD domain leads to the loss of stem cell phenotypes and downregulation of the FGF signaling. Moreover, impairment of RING or PHD domain results in decreased proliferation and impedes embryoid body formation. In opposition to previous data indicating the impact of phosphorylation on TRIM28 function, our data suggest that TRIM28 phosphorylation does not significantly affect the pluripotency and self-renewal maintenance of hiPSC. Of note, iPSC with disrupted RING and PHD functions display downregulation of genes associated with tumor metastasis, which are considered important targets in cancer treatment. Our data suggest the potential use of RING and PHD domains of TRIM28 as targets in cancer therapy.

Sign in / Sign up

Export Citation Format

Share Document