Isolating Cells and Growing them in Culture

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What is the first step in isolating cells of a uniform type from a tissue?

Disrupting the extracellular matrix

What type of tissues usually yield the best dissociated cells?

Fetal or neonatal tissues

What type of enzymes are typically used to digest proteins in the extracellular matrix?

Proteolytic enzymes like trypsin and collagenase

What is the role of ethylenediaminetetraacetic acid (EDTA) in isolating cells?

Binding or chelating Ca2+ on which cell-cell adhesion depends Signup and view all the answers

How can large cells be separated from small cells in a mixed cell suspension?

By centrifugation Signup and view all the answers

What is the purpose of gentle agitation in isolating cells from tissue?

To tease the tissue into single living cells Signup and view all the answers

What is the purpose of isolating cells from a tissue and growing them in culture?

To obtain a relatively homogenous population of cells for detailed biochemical analysis. Signup and view all the answers

Why do most biochemical procedures require obtaining large numbers of cells?

To physically disrupt the cells and isolate their components for analysis. Signup and view all the answers

Why do biologists develop ways to dissociate cells from tissues and separate various types?

To obtain as much information as possible about individual cell types. Signup and view all the answers

What is the benefit of allowing isolated cells to proliferate as a pure culture?

To greatly increase the number of cells for analysis. Signup and view all the answers

How can heterogenous cell populations in a tissue be handled to analyze individual cell types?

By dissociating cells from tissues and separating the various types. Signup and view all the answers

What is the significance of obtaining a homogenous population of cells for analysis?

It allows for more accurate and specific study of cell components. Signup and view all the answers

How can cells be separated based on their tendency to adhere strongly to glass or plastic?

Cells can be separated based on their tendency to adhere strongly to glass or plastic by allowing them to adhere and then separating them from cells that adhere less strongly. Signup and view all the answers

What is the principle behind separating cells by antibodies coupled to various matrices?

Antibodies that bind specifically to the surface of only one cell type in a tissue are coupled to matrices to create an affinity surface for cell adhesion. Signup and view all the answers

How are cells recovered after being bound to antibodies coupled to matrices?

Cells bound to antibodies coupled to matrices can be recovered by gentle shaking, treatment with trypsin, or degrading the matrix itself with enzymes. Signup and view all the answers

What are some examples of matrices that can be used along with antibodies for cell separation?

Collagen, polysaccharide beads, or plastic can be used as matrices along with antibodies for cell separation. Signup and view all the answers

Explain how trypsin can be used in the process of recovering cells bound to antibodies.

Trypsin can be used to digest the proteins that mediate the adhesion of cells bound to antibodies, facilitating their release. Signup and view all the answers

What is the role of collagenase in cell separation using digestible matrices?

Collagenase degrades the digestible matrix, such as collagen, to release cells bound to antibodies for separation. Signup and view all the answers

Isolating Cells and Growing them in Culture - BSC302 (Lecture 10)

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