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hiPS Cell Biology Overview

Insights about hIPS cells & what we have learned by growing, manipulating & imaging them.
Back to About our Cells
 

Summary on the epithelial nature of hiPS cells grown on 2D substrate​

Picture
We have a series of observations based on the localization pattern of 10 intracellular structures that are all consistent with an apical-basal polarity and epithelial sheet nature of hiPS cell colonies:
  • An apical-basal polarity of microtubules, which may also lead to differences in apical vs. basal density of other cytoplasmic organelles (mitochondria, ER, Golgi).
  • An apical band of bundled actin, myosin, tight junctions, and desmosomes around the apical region of the cell.
  • Localization of actin-containing structures at the basal surface of the cells.
  • The apical localization of centrioles and primary cilia.
  • View these features on the Cell Structure Observations page.

hiPS cells grow in single-cell layer colonies on 2D surfaces​

Bright field movie of entire colony in time 
Close up of colony (division occurring near top)​
Figure 1. Transmitted light timelapse movies of live hiPSC cells imaged on a spinning-disk confocal microscope. ​Images were collected every 5 minutes for 3 hours. Playback speed is 3000x real time. 
​Observations
  • Colonies are very dynamic, with cells growing, dividing, and dying (turn-over) within the colony as the colony itself grows and expands. The colony edges are constantly protruding, extending lamellipodia and retracting. Cells within healthy colonies are tightly packed next to each other in a single sheet.
  • Cells are approximately 10-20 microns tall and 5-20 microns in diameter. Over time the cells at the center of colonies become extremely tightly packed and flatten. As the cell density in this region increases, more cells undergo apoptosis and are expelled upward out of the monolayer. Cells at the edges of colonies are much flatter, with large flat lamellipodial regions. Cells between the center and edge of a colony are the tallest, forming a ‘ridge’.
  • As cells move, they push and pull on each other, exhibiting a coherence in their movement. Dividing cells usually divide near the top of the colony, and then intercalate back into the colony, but often not directly adjacent to each other.
  • Groups of cells residing near each other often seem to divide at about the same time (see also e.g. timelapse movies of colonies of cells expressing tagged α-tubulin below). This could be because sister cells may be more in synchrony with each other and also closer to each other within the colony.

General organization of hiPS cells​

A. Microtubules
B. Mitochondria
C. Golgi
Picture
D. 3D colony rendering with interpreted cell illustration
Figure 2. Z-stack movies of live hiPS cells. Three examples of cells expressing mEGFP tagged α-tubulin (A) in microtubules, Tom20 (B) in mitochondria, and ß-galactoside α-2,6-sialyltransferase 1 (C) in Golgi imaged on a spinning-disk confocal microscope. Images start from the bottom of the cells and end at the top. The left and center movie include fluorescent dyes that label the cell membrane (magenta) and the DNA/nucleus (cyan). (D) 3D illustration of the organization of hiPS cells in a monolayer colony.
​Observations
  • In general, cells are columnar with differences in the distribution of intracellular structures and organelles depending on their apical-basal location. However, the cells often lean over each other such that they are not necessarily cylindrical but instead can form a wide range of 3D shapes (see Modeling & Analysis section of the website).
  • The nucleus in these cells occupies ~30-50% of the cell volume and most of the cytoplasmic space especially in the center region of the cells.
  • Key organelles and structures localize more densely in a cytoplasmic ‘pocket’ above the nucleus and are sparser towards the bottom of the cell. Example structures that exhibit this localization pattern include microtubules, which seem to emanate from the top to the bottom (see microtubule section), mitochondria, which are more densely clustered in the top, and the Golgi which is primarily localized to the top of cells.

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  • About
      Institute
      1. News feed
      2. What we do
      3. Publications
      4. Allen Institute | allenInstitute.org
      5. Careers
      Site
      1. Home page
      2. Site updates
      3. Archived content
  • Allen Cell Collection
      Order cells & plasmids
      1. Cell Catalog
      2. Cell Catalog quickview
      3. Cell Shorts (documentaries on labs using our cells)
      4. Support forum
      Lab methods
      1. Instructional videos for success in the lab
      2. Standard operating procedures (written methods)
      3. Illustrated overviews
      About our hiPS cells
      1. hiPS Cell Structure Overview
      2. Visual Guide to Human Cells
      3. Cell structure observations
      4. Why endogenous tagging?
      5. Differentiation into cardiomyocytes
      6. Genomics
      7. Download cell data (images, genomics, features)
  • Data & Digital Tools
      Online image analysis
      1. Cell Feature Explorer (plotting & 3D viewer)
      2. 3D cell viewer (pre2018)
      3. Deep cell zoom (216,016 cells)
      Online modeling viewers
      1. Visual Guide to Human Cells
      2. Simularium (4D visual analysis)
      3. Integrated Mitotic Stem Cell
      4. └ Z-stack viewer
      5. └ 3D viewer
      6. Allen Integrated Cell viewer
      7. Label-free examples viewer
      8. 3D probabilistic model viewer
      Desktop tools
      1. Allen Cell & Structure Segmenter
      2. AGAVE 3D pathtrace image viewer
      Data & code
      1. Download cell data (images, genomics, features)
      2. Code repositories & software
  • Analysis & Modeling
      Allen Integrated Cell models
      1. Overview
      2. Integrated Mitotic Stem Cell
      3. └ Z-stack viewer
      4. └ 3D viewer
      5. Label-free Determination
      6. └ 3D viewer
      7. 3D Probabilistic Modeling
      8. └ 3D viewer
      9. Visual Guide to Human Cells
      4D biology models
      1. Simularium (online 4D viewer)
      Methodologies
      1. Drug perturbation pilot study
      2. hiPS cells during mitosis
      3. Differentiation into cardiomyocytes
  • Publications
      Articles
      1. All journal publications
      2. Preprints (biorxiv, arxiv)
      Posters
      1. Select posters
  • Education
      Education resources
      1. All Resources
      2. Teaching materials
      Online tools popular with teachers
      1. Visual Guide to Human Cells
      2. Integrated Mitotic Stem Cell
      3. Cell Feature Explorer (interactive plotting & 3D viewer)
      4. 3D cell viewer (pre2018 data)
      5. hiPS cell structure overview
  • Support
      Questions
      1. FAQs
      2. Forum
      Tutorials for digital tools
      1. Digital tool tutorials with videos
      2. Visual Guide tutorial
      3. AGAVE user guide
      Lab methods
      1. Instructional videos for success in the lab
      2. Standard operating procedures (written methods)
      3. Illustrated overviews
  • 🔍
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