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Isolation and Identification of Limbal Niche Cells

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10:11 min

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October 27th, 2023

DOI :

10.3791/65618-v

October 27th, 2023

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Guanyu Su¹, Wei Wang¹, Lingjuan Xu¹, Rong Liu¹, Yongyao Tan¹, Bihui Jin¹, Chunxiu You¹, Xiao Zhou¹, Yihong Xiong², Huatao Xie³, Guigang Li¹

¹Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, ²Institute of Medicine Nursing, Hubei University of Medicine, ³Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology

Transcript

Our studies focus on the reconstruction of the limbal niche to treat limbal stem cell deficiency and to restore the patient's vision. We are trying to answer whether or not limbal niche cells are the key cells supporting the function of limbal stem cells, and if so, how it works. Recent studies reported that subconjunctival transplantation of mesenchymal stem cells from bone marrow prevented the limbal stem cell deficiency of alkali-burned eyes.

One of the technologies currently used to advance our research is single cell sequencing, which can analyze the key signaling pathway activities at a single cell level for limbal niche cell characterizing. Although LNC have the potential to become an important clinical tool in the future, their growth characteristics have not been explored in much details. Considering their promising scientific and clinical applications, it is necessary to summarize the isolation, purification, identification, and characteristics of LNC.

Summary

Here, we present a protocol to isolate and identify the human limbal niche cells.

Explore More Videos

Limbal Stem Cell Deficiency

Single Cell Sequencing

Purification

Cell Culture

Immunofluorescence

Real time Quantitative PCR

Flow Cytometry

Stem Cell Markers

Cell Proliferation

Mesenchymal Stem Cells BMMSCs

Tissue Transplantation

Chapters in this video

0:00

Introduction

1:25

Isolation of Limbal Niche Cells (LNCs) from Limbus Tissue