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Virginia Tech

Engineering Organoids for Stem Cell Maturation

Abstract

dc:description.abstractgeneral

The liver is the largest internal organ in the body. It is responsible for performing an array of vital functions, including the filtration of blood, synthesis of different molecules, and metabolism of drugs and toxicants. Hepatocytes, or the main liver cell type, perform most of these functions. Typically, primary human hepatocytes (PHHs) are ideal for in vitro liver studies since they are obtained directly from tissues and exhibit adult characteristics and functions. However, sourcing these adult liver cells is extremely difficult since they are usually obtained through biopsies and are limited in quantity. Induced pluripotent stem cell (iPSC)-hepatocyte-like cells (iHLCs) hold tremendous potential to be used as a substitute since they can be obtained non-invasively. However, iHLCs require further maturation before they can be substituted for PHHs due to their lower liver-related functions and immature characteristics. Existing maturation approaches require the administration of chemical mixtures that can be up to a million times higher than the concentrations of these same molecules inside the body. The lack of a systematic approach to mature iHLCs currently limits their widespread use. During liver development, neighboring cells secrete different proteins that regulate and induce multiple pathways that aid in the maturation of cells into adult liver cells. We report the assembly of a multicellular 3D human liver organoid with iHLCs that recapitulates the in vivo hepatic microenvironment. Intra- and intercellular signaling between human hepatic cells in the organoid result in mature iHLCs that exhibit several markers and functions of PHHs within one to two weeks in culture. When two other hepatic cell types, Kupffer cells and liver sinusoidal endothelial cells, are present in the organoids, they secrete signaling molecules that synergistically mature iHLCs. Relying solely on intercellular secretion from the cells provides a systematic and reproducible approach to generate mature iHLCs for drug, disease, and patient-specific in vitro studies.

Degree

thesis:*
Name thesis:degree_name
Doctor of Philosophy
Level thesis:degree_level
doctoral
Discipline thesis:degree_discipline
Chemical Engineering
Department dc:contributor.department
Chemical Engineering
Grantor dc:publisher
Virginia Tech
Year dc:date.issued
2024

Author and committee

dc:creator, dc:contributor.*
Author dc:creator
  • Gandhi, Neeti Nimish
Chair dc:contributor.committeechair
  • Rajagopalan, Padmavathy
Committee members dc:contributor.committeemember
  • Wrenn, Steven Parker
  • Lee, Yong Woo
  • Tong, Rong
  • Davis, Richey M.

Subjects

dc:subject × 5

Rights

dc:rights
Statement dc:rights
  • In Copyright
Language dc:language.iso
en

Identifiers

dc:identifier.*
Dc Identifier Other
vt_gsexam:41770
OAI identifier oai:identifier
oai:vtechworks.lib.vt.edu:10919/123753

Chain of custody

source
Harvested from
Virginia Tech
Base URL
vtechworks.lib.vt.edu/oai/request
Last updated
2026-07-22
Source record
OAI-PMH GetRecord
citation

Gandhi, Neeti Nimish. Engineering Organoids for Stem Cell Maturation. doctoral thesis, Virginia Tech, 2024. https://hdl.handle.net/10919/123753