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hiPS cell structure overview

Insights about hIPS cells & what we have learned by growing, manipulating & imaging them

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​

Microtubules
Mitochondria
Golgi
Picture
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.
  • 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. Our science: CellScapes
      2. Past foundational projects
      3. News feed
      4. About us
      5. Careers
  • Allen Cell Collection
      Order cells & plasmids
      1. Cell Catalog
      2. Disease Collection Cell Catalog
      3. Cell Catalog quickview
      4. Cell video shorts
      Lab methods
      1. Video protocols
      2. Written protocols
      3. Our methodology
      4. Support forum
      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
      General
      1. Tools and resources overview
      2. Download cell data (images, genomics, features)
      3. Code repositories & software
      Desktop tools
      1. Allen Cell & Structure Segmenter
      2. AGAVE 3D pathtrace image viewer
      Web tools
      1. BioFile Finder
      2. Cell Feature Explorer
      3. Integrated Mitotic Stem Cell
      4. └ Z-stack viewer
      5. └ 3D viewer
      Web tools (con't)
      1. Simularium viewer
      2. Timelapse Feature Explorer
      3. Visual Guide to Human Cells
      4. Vol-E (Web Volume Viewer)
      5. 3D Cell Viewer
  • Analysis & Modeling
      Allen Integrated Cell models
      1. Visual Guide to Human Cells
      2. Integrated Mitotic Stem Cell
      3. └ Z-stack viewer
      4. └ 3D viewer
      5. Allen Integrated Cell
      6. └ 3D Probabilistic Modeling
      7. └ Label-free Determination
      4D biology models
      1. Simularium viewer
      Methodologies
      1. Drug perturbation pilot study
      2. hiPS cells during mitosis
      3. Differentiation into cardiomyocytes
  • Publications
      Articles
      1. Publications
      2. Preprints
      Presentations
      1. Talks & posters
  • Education
      Educational resources
      1. All resources
      2. Teaching materials
      Online tools popular with teachers
      1. Visual Guide to Human Cells
      2. Integrated Mitotic Stem Cell
      3. 3D Cell Feature Explorer
      4. 3D Cell Viewer
      5. hiPS cell structure overview
  • Support
      Questions
      1. FAQs
      2. Forum
      Tutorials for digital tools
      1. Video tutorials
      2. Visual Guide tutorial
      3. AGAVE documentation
      Lab methods
      1. Video protocols
      2. Written protocols
      3. Our methodology
  • 🔍
      SEARCHBAR