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JoVE Journal
Immunology and Infection
Salispheres veya Monolayers olarak In vitro büyüme için ınsan tükürük bezlerinden tükürük epitely...
Salispheres veya Monolayers olarak In vitro büyüme için ınsan tükürük bezlerinden tükürük epitely...
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Isolation of Salivary Epithelial Cells from Human Salivary Glands for In Vitro Growth as Salispheres or Monolayers

Salispheres veya Monolayers olarak In vitro büyüme için ınsan tükürük bezlerinden tükürük epitelyal hücrelerin yalıtım

Full Text
9,764 Views
08:03 min
July 15, 2019

DOI: 10.3791/59868-v

Matthew J. Beucler1, William E. Miller1

1Department of Molecular Genetics, Biochemistry, and Microbiology,University of Cincinnati College of Medicine

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Overview

This article presents a method for isolating and cultivating primary human salivary gland-derived epithelial cells. These cells can be grown as salispheres or monolayers, providing a more in vivo-like model for studying human viruses.

Key Study Components

Area of Science

  • Cell Biology
  • Virology
  • Regenerative Medicine

Background

  • Primary cellular models are essential for biological research.
  • Salivary glands play a crucial role in viral transmission.
  • Existing cell models often do not accurately represent in vivo conditions.
  • This study focuses on developing a new protocol for salivary gland cells.

Purpose of Study

  • To isolate and cultivate primary human salivary gland-derived epithelial cells.
  • To create a model that mimics in vivo conditions for studying viral replication.
  • To provide a protocol applicable to various patient samples.

Methods Used

  • Isolation of epithelial cells from human salivary glands.
  • Culture of cells as salispheres on basement membrane matrices.
  • Growth of cells as monolayers on treated culture dishes.
  • Application of the protocol to mouse primary submandibular gland cells.

Main Results

  • Successful isolation of primary salivary gland-derived epithelial cells.
  • Cells exhibited gene expression patterns consistent with salivary epithelial origin.
  • Protocol allows for the generation of cells from various patient samples.
  • Potential applicability to other organs with similar cellular structures.

Conclusions

  • The developed protocol is a significant advancement in cellular modeling.
  • It enhances the understanding of viral mechanisms in salivary glands.
  • This method could facilitate future research on organ-specific viral interactions.

Frequently Asked Questions

What are salispheres?
Salispheres are three-dimensional structures formed by cultured salivary gland-derived epithelial cells, resembling in vivo tissue.
Why is studying salivary glands important?
Salivary glands are key sites for viral replication and transmission, making them crucial for understanding viral diseases.
Can this protocol be used for other organs?
Yes, the protocol may be applicable to other organs with similar cellular structures.
What is the significance of using primary cells?
Primary cells provide a more accurate representation of in vivo conditions compared to transformed cell lines.
How does this method improve upon existing techniques?
This method allows for the generation of more physiologically relevant cellular models from patient samples.
What are the potential applications of this research?
The research could lead to better understanding of viral mechanisms and development of therapeutic strategies.

Birincil insan tükürük bezi türevi epitelyal hücreleri izole etmek ve yetiştirmek için bir yöntem sunuyoruz. Bu hücreler onları tükürük epitelyal orijinli olmak ile tutarlı gen ifade desenleri sergiler ve Engelbreth-Holm-Swarm tümör hücrelerinden türetilen Bodrum membran matrisleri üzerinde salispheres olarak veya tedavi kültür yemekleri monolayers olarak yetişebilir.

Birincil hücresel modeller veya organoid benzeri yapılar üretmek biyolojideki birçok güncel soruyu yanıtlamanın anahtarıdır. Örneğin, yatay iletim için önemli bir site olduğu gibi tükürük bezi çoğaltma kolaylaştırmak için insan virüsleri tarafından kullanılan mekanizmalar oldukça uzun bir süre ilgi oldum. Bu protokolde tanımlanan salya temelli sistemin gelişimi, bu tür sorulara cevap verme arayışımızda çok önemli bir ilerlemedir.

Bu teknik bize farklı hasta örnekleri çeşitli birincil hücreleri oluşturmak için izin verir. Hücreler in vitro deneyler için yaygın olarak kullanılan birçok hücre tipi gibi modellenmiş veya dönüştürülmüş değil gibi hücresel modeller in vivo tipi durumu daha anımsatır. Bu protokolü fare nin birincil submandibular bezinden benzer hücreler üretmek için kullandık ve protokol benzer hücresel yapılara sahip diğer organlar için de yararlı olacaktır.

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