Comparison of Two Representative Methods for Differentiation of Human Induced Pluripotent Stem Cells into Mesenchymal Stromal Cells

Fangqin Wang; Langping Gao; Xudong Fu; Qintao Yan; Lidan Hu; Jianhua Mao

doi:10.3791/65729

İnsan Kaynaklı Pluripotent Kök Hücrelerin Mezenkimal Stromal Hücrelere Farklılaşması için İki Tem...

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Comparison of Two Representative Methods for Differentiation of Human Induced Pluripotent Stem Cells into Mesenchymal Stromal Cells

İnsan Kaynaklı Pluripotent Kök Hücrelerin Mezenkimal Stromal Hücrelere Farklılaşması için İki Temsili Yöntemin Karşılaştırılması

Full Text

1,768 Views

06:24 min

October 20, 2023

DOI: 10.3791/65729-v

Fangqin Wang¹, Langping Gao^2,3, Xudong Fu⁴, Qintao Yan², Lidan Hu^1,2, Jianhua Mao^2,3

¹National Clinical Research Center for Child Health,The Children's Hospital, Zhejiang University School of Medicine, ²Department of Nephrology,The Children's Hospital, Zhejiang University School of Medicine, ³Zhejiang University School of Medicine, ⁴Liangzhu Laboratory,Zhejiang University

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study explores the differentiation of human induced pluripotent stem cells (hiPSCs) into mesenchymal stromal cells (MSCs) using two distinct methods: the monolayer method and the embryoid bodies (EBs) method. The monolayer method offers cost-effectiveness and simplicity, particularly in osteogenic differentiation, while the EBs method is noted for its reduced time requirements.

Key Study Components

Research Area

Stem cell differentiation
Mesenchymal stromal cells
Human induced pluripotent stem cells

Background

Importance of directed differentiation of iPSCs
Challenges in generating specific cell types from stem cells
Applications in genetic disease models

Methods Used

Monolayer and EBs differentiation methods
Human induced pluripotent stem cells
Flow cytometry for cell characterization

Main Results

Monolayer-derived MSCs showed more calcium deposits compared to EBs-derived MSCs.
Both methods demonstrated similar abilities in adipogenic and chondrogenic differentiation.
hiPSCs maintained proliferation abilities across multiple passages.

Conclusions

The study highlights the effectiveness of both differentiation methods for generating MSCs.
Findings contribute to the understanding and application of stem cell therapies in genetic disorders.

Frequently Asked Questions

What are mesenchymal stromal cells?

Mesenchymal stromal cells (MSCs) are multipotent stem cells that can differentiate into a variety of cell types, including bone, cartilage, and fat cells.

Why is differentiation of hiPSCs important?

Differentiating hiPSCs into specific cell types is crucial for modeling diseases and developing potential therapies.

What advantages do hiPSCs offer in research?

hiPSCs share the same genome as the patient, providing an accurate model for studying genetic diseases.

How do you measure the success of MSC differentiation?

Success is often measured by the expression of specific surface markers and the cells' ability to differentiate into various lineages.

What are the common applications of MSCs?

MSCs are used in regenerative medicine, tissue engineering, and as models for studying disease mechanisms.

What is the role of flow cytometry in this study?

Flow cytometry is used to characterize the surface markers of differentiated cells to confirm their identity as MSCs.

Can both differentiation methods be used interchangeably?

While both methods yield MSCs, their efficiency and specific characteristics might vary, making them suitable for different applications.

Bu protokol, hiPSC'leri mezenkimal stromal hücrelere (MSC'ler) farklılaştırmak için iki temsili yöntemi tanımlar ve karşılaştırır. Tek katmanlı yöntem, daha düşük maliyet, daha basit işlem ve daha kolay osteojenik farklılaşma ile karakterizedir. Embriyoid cisimler (EB'ler) yöntemi, daha düşük zaman tüketimi ile karakterizedir.

Çalışmamızın kapsamı, genetik hastalıkların bir mekanizması ve terapötik strateji araştırmasıdır. Araştırmamız, hücre hatları, iPSC'ler, organoidler ve fareler gibi çok düzeyli hastalık modellerine dayanmaktadır. iPSC'lerin diğer modellere göre belirgin bir avantajı vardır.

Hastalardan alınan iPSC'ler ve daha sonra türetilen hücreler ve organoidler, hastalarla aynı genomu paylaşır ve bu da birçok hastalık için bir hasta hücresi ve model bankası sağlar. Ayrıca, hastalardan kurulan iPSC'lerin yöntemleri ile oldukça olgunlaşmıştır. iPSC'lerin spesifik hücrelere ve organoidlere farklılaşması yönünde birçok zorluk devam etmektedir ve mevcut gelişimde geliştirme mekanizmalarının derinleşmesi alanında ortaya çıkacaktır.

View the full transcript and gain access to thousands of scientific videos

Explore More Videos

Mezenkimal Stromal Hücreler MKH'ler Rejeneratif Tıp İnsan Kaynaklı Pluripotent Kök Hücreler HiPSC'ler Farklılaşma Kültür Metodolojileri Embriyoid Cisimler Tek Tabakalı Kültür Avantajları Dezavantajları Zaman Tüketimi Maliyet Hücre Proliferasyon Yeteneği MSC Belirteçleri İn Vitro Farklılaşma Yeteneği Matür MSC'ler