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A Two-Step Strategy that Combines Epigenetic Modification and Biomechanical Cues to Generate Mammalian Pluripotent Cells

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A process engineering approach to increase organoid yield.

Natasha Arora¹, Jasmin Imran Alsous², Jacob W Guggenheim³

¹Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA.

Development (Cambridge, England)

|February 9, 2017

Summary

This summary is machine-generated.

Researchers identified key features of early-stage intestinal organoids (spheroids) to improve their generation. Larger spheroids with an inner mass are more likely to develop into high-quality intestinal organoids.

Keywords:

Aggregates Intestinal Organoids Process engineering Sorting Yield

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Area of Science:

Stem cell biology
Developmental biology
Tissue engineering

Background:

Human pluripotent stem cells (hPSCs) can differentiate into organoids, mimicking native organs.
Understanding early organoid formation is crucial for robust protocols in regenerative medicine and drug development.

Purpose of the Study:

To investigate the emergence of intestinal organoids from hindgut spheroids.
To identify measurable parameters predicting successful organoid maturation.

Main Methods:

Focused on hindgut spheroids as an intermediate step in intestinal organoid development.
Utilized an automated micropipette system to sort spheroids based on diameter and morphology.
Monitored sorted spheroid cultures for organoid formation.

Main Results:

13% of spheroids successfully matured into intestinal organoids.
Spheroid diameter and the presence of an inner mass were identified as key maturation parameters.
Spheroids >75 μm diameter were 1.5-fold enriched for pre-organoids.
Spheroids with an inner mass were 3.8-fold enriched for pre-organoids.

Conclusions:

Diameter and inner mass are critical indicators for predicting intestinal organoid maturation.
These findings provide guidelines for developing robust protocols for high-quality intestinal organoid generation.