# A bio-engineered hepatic niche for ex vivo expansion of HSCs

> **NIH NIH R01** · UNIVERSITY OF ILLINOIS AT CHICAGO · 2023 · $319,800

## Abstract

Abstract / Project Summary
New Directions in Hematology Research (SHINE-II) PAS-18-730 NIDDK
Title: Expansion of bone marrow hematopoietic stem cells in modular human liver culture platforms
The hematopoietic stem cell (HSC) resides at the top of the hierarchy of blood cells and is responsible for the
generation of all the progenitor and mature cells of the hematopoietic system. How HSCs are capable of
generating the vast diversity of hematopoietic cell types while still retaining their self-renewal and proliferative
capacity is a topic of active research in the field. Many studies have pointed to the niche as the critical source of
extrinsic factors that instruct and support HSC cell fate decisions. The primary niche for homeostatic
hematopoiesis in adult mammals is the bone marrow. Much debate still exists regarding the specific cell types
of the bone marrow niche, but it is generally accepted that in the bone marrow HSCs are primarily quiescent and
infrequently activate for expansion and differentiation of progenitor cell types. Likewise, experiments that have
attempted to expand HSCs ex vivo using bone marrow stromal cells or cytokine cocktails in tissue culture plastic
have not been successful in generating robust and renewable stem cells capable of long-term, multi-lineage
reconstitution of irradiated recipients. We hypothesize that an approach based on the ontogeny of hematopoietic
system will allow for a more successful ex vivo expansion of HSCs. Previous findings have shown that HSCs
expand very rapidly during embryonic development in the fetal liver. Furthermore, under stress or due to certain
pathological conditions, HSCs migrate to the adult liver where they undergo extramedullary hematopoiesis
(EmH). Following these physiological cues, we propose to bioengineer ex vivo co-culture systems of increasing
complexities using primary human hepatocytes (PHH), supportive fibroblasts, and primary human liver non-
parenchymal cells (NPC). We will first utilize our long-term micropatterned co-culture (MPCC) system of PHH
colonies surrounded by supportive fibroblasts (3T3-J2 murine embryonic fibroblasts and human liver portal
fibroblasts or LPF) to which we will introduce adult bone marrow HSCs cultured in an optimized serum-free
culture medium supplemented with hematopoietic cytokines. We will monitor HSC expansion in MPCCs and
characterize them phenotypically via flow cytometry prior to assessing their functional potential in vitro and in
vivo in mouse transplantation assays. Next, we will incorporate HSCs into novel 3-dimensional (3D) human liver
microtissues that we have shown to enable optimal interactions of PHHs and primary human liver sinusoidal
endothelial cells (LSEC); the PHH-LPF-LSEC microtissues will be further augmented with quiescent hepatic
stellate cells and Kupffer cells to elucidate the role of these cell types in modulating HSC expansion ex vivo.
Lastly, we will induce EmH in mice to prime HSCs for the liver microenvironment ...

## Key facts

- **NIH application ID:** 10631071
- **Project number:** 5R01HL151720-03
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT CHICAGO
- **Principal Investigator:** Salman R Khetani
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $319,800
- **Award type:** 5
- **Project period:** 2021-07-16 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10631071

## Citation

> US National Institutes of Health, RePORTER application 10631071, A bio-engineered hepatic niche for ex vivo expansion of HSCs (5R01HL151720-03). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10631071. Licensed CC0.

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