Quantifying the fidelity of in vitro human cell culture systems using a biomedical foundation model

S Satoru Fujii (Department of Gastroenterology and Hepatology, Institute of Science Tokyo) S Scott T. Espenschied (Department of Inflammation and Immunity, Cleveland Clinic Research, The Cleveland Clinic) V Vibha Anand (Healthcare and Life Sciences, International Business Machines Corporation Research) J Joao Bettencourt-Silva (Healthcare and Life Sciences, International Business Machines Corporation Research) Y Yi Han G Go Ito (Department of Gastroenterology and Hepatology, Institute of Science Tokyo) A Akira Koseki (Healthcare and Life Sciences, International Business Machines Corporation Research) A Akihiro Kosugi (Healthcare and Life Sciences, International Business Machines Corporation Research) J James Kozloski (Healthcare and Life Sciences, International Business Machines Corporation Research) R Ryoma Matsumoto (Department of Gastroenterology and Hepatology, Institute of Science Tokyo) S Shanshan Meng N Natasha Mulligan (Healthcare and Life Sciences, International Business Machines Corporation Research) R Ryan J. Musich (Department of Inflammation and Immunity, Cleveland Clinic Research, The Cleveland Clinic) K Kevin P. Newhall (Department of Inflammation and Immunity, Cleveland Clinic Research, The Cleveland Clinic) E Eri Oshina (Department of Gastroenterology and Hepatology, Institute of Science Tokyo) S Shuhei Sekiguchi (Department of Gastroenterology and Hepatology, Institute of Science Tokyo) Y Yi Wang J Jianying Hu (Healthcare and Life Sciences, International Business Machines Corporation Research) M Matthew Ciorba (Division of Gastroenterology and the Inflammatory Bowel Diseases Center, Department of Internal Medicine, Washington University School of Medicine) L L. David Sibley R Ryuichi Okamoto (Department of Gastroenterology and Hepatology, Institute of Science Tokyo) T Thaddeus S. Stappenbeck (Department of Inflammation and Immunity, Cleveland Clinic Research, The Cleveland Clinic)

Abstract

Primary cell culture is fast becoming a dominant method for discovery work regarding human disease. Currently, there are no methods to quantitatively benchmark these systems. Here, we apply a uniform in vitro culture system of human intestinal epithelial cells (IECs) to achieve this goal. We previously established methods for long-term two-dimensional (2D) cultivation of mouse IECs using an air–liquid interface (ALI) technique. Here, we further refined these methods for long-term 2D cultivation of human IECs, with histological and molecular features of differentiated intestinal epithelia. Leveraging the power and scalability of a biomedical foundation model (BMFM) trained on single-cell RNA sequencing data (BMFM-RNA), we performed classification tasks to identify cell types across sample sources and to quantitatively benchmark our in vitro differentiated cells against cells collected from patient biopsies. We observed a striking concordance between our in vitro differentiated cells and the corresponding cell types in vivo for multiple differentiated secretory cell types. This approach using BMFM-RNA holds promise to expand our understanding of the regulatory mechanisms, including gene–gene regulation underlying homeostasis and regeneration, as well as the functions of rare and poorly understood lineages within the human intestinal epithelia. Moreover, these methods may be applicable to other organs, model systems, and experimental modalities. We propose that the framework used here can be deployed as a standard benchmarking methodology, ultimately improving the fidelity of primary human culture systems.

Article Details

Volume / Issue Vol. 123, Issue 12
Published March 24, 2026
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (22)

S

Satoru Fujii

Department of Gastroenterology and Hepatology, Institute of Science Tokyo

S

Scott T. Espenschied

Department of Inflammation and Immunity, Cleveland Clinic Research, The Cleveland Clinic

V

Vibha Anand

Healthcare and Life Sciences, International Business Machines Corporation Research

J

Joao Bettencourt-Silva

Healthcare and Life Sciences, International Business Machines Corporation Research

Y

Yi Han

G

Go Ito

Department of Gastroenterology and Hepatology, Institute of Science Tokyo

A

Akira Koseki

Healthcare and Life Sciences, International Business Machines Corporation Research

A

Akihiro Kosugi

Healthcare and Life Sciences, International Business Machines Corporation Research

J

James Kozloski

Healthcare and Life Sciences, International Business Machines Corporation Research

R

Ryoma Matsumoto

Department of Gastroenterology and Hepatology, Institute of Science Tokyo

S

Shanshan Meng

N

Natasha Mulligan

Healthcare and Life Sciences, International Business Machines Corporation Research

R

Ryan J. Musich

Department of Inflammation and Immunity, Cleveland Clinic Research, The Cleveland Clinic

K

Kevin P. Newhall

Department of Inflammation and Immunity, Cleveland Clinic Research, The Cleveland Clinic

E

Eri Oshina

Department of Gastroenterology and Hepatology, Institute of Science Tokyo

S

Shuhei Sekiguchi

Department of Gastroenterology and Hepatology, Institute of Science Tokyo

Y

Yi Wang

J

Jianying Hu

Healthcare and Life Sciences, International Business Machines Corporation Research

M

Matthew Ciorba

Division of Gastroenterology and the Inflammatory Bowel Diseases Center, Department of Internal Medicine, Washington University School of Medicine

L

L. David Sibley

R

Ryuichi Okamoto

Department of Gastroenterology and Hepatology, Institute of Science Tokyo

T

Thaddeus S. Stappenbeck

Department of Inflammation and Immunity, Cleveland Clinic Research, The Cleveland Clinic