Isolation of Salivary Epithelial Cells from Human Salivary Glands for In Vitro Growth as Salispheres or Monolayers

Matthew  J. Beucler; William E. Miller

doi:10.3791/59868

Isolamento delle cellule epiteliali salivari dalle ghiandole salivari umane per la crescita in vi...

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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

Isolamento delle cellule epiteliali salivari dalle ghiandole salivari umane per la crescita in vitro come Salispheres o Monostrati

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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.

Presentiamo un metodo per isolare e coltivare cellule epiteliali derivate dalla ghiandola umana primaria. Queste cellule presentano modelli di espressione genica coerenti con loro di origine epiteliale salivari e possono essere coltivate come salisfere su matrici di membrana seminterrato derivate da cellule tumorali Engelbreth-Holm-Swarm o come monostrati su piatti di coltura trattati.

Generare modelli cellulari primari o strutture organoidi è la chiave per rispondere a molte domande contemporanee in biologia. Ad esempio, siamo interessati da un bel po 'di tempo ai meccanismi utilizzati dai virus umani per facilitare la replicazione nella ghiandola salivare in quanto è un sito importante per la trasmissione orizzontale. Lo sviluppo del sistema basato sulla salisfera descritto in questo protocollo è un progresso cruciale nella nostra ricerca di rispondere a questo tipo di domande.

Questa tecnica ci consente di generare cellule primarie da una varietà di diversi campioni di pazienti. I modelli cellulari ricordano più la situazione di tipo in vivo in quanto le cellule non sono modellate o trasformate come molti tipi di cellule comunemente usati per esperimenti in vitro. Abbiamo usato questo protocollo per generare cellule simili dalla ghiandola submandibolare primaria del topo e il protocollo sarebbe probabilmente utile per altri organi con strutture cellulari simili.

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