Pioneers of Tissue Culture

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Questions and Answers

Which of the following best describes the lasting impact of Kohler and Milstein's 1975 work?

Improved methods for long-term cell line maintenance.
Enabled the large-scale production of precisely targeted vaccines. (correct)
Advanced the use of in vitro fertilization techniques.
Facilitated the study of early embryonic development.

What key challenge did Steptoe and Edwards overcome in 1977 that led to a major reproductive breakthrough?

Developing methods for monoclonal antibody production.
Maintaining neural tissues ex vivo indefinitely.
Advancing microscopic evaluation techniques for cell lines.
Successfully culturing early embryos in vitro. (correct)

What was Wilhelm Roux's primary contribution to the advancement of tissue culture techniques?

He maintained neural folds from early chick embryos in saline solution. (correct)
He embedded frog embryos in blood clots for microscopic evaluation.
He developed in vitro fertilization techniques.
He was the first to generate a continuous cell line from embryonic chicken heart tissue.

How did Ross Granville Harrison contribute to the field of tissue culture?

By embedding small sections of frog embryos in blood clots to allow microscopic evaluation. (D) Signup and view all the answers

What was the significance of Alexis Carrel and Montrose Thomas Burrows's contribution to tissue culture in 1911?

They generated the first continuous cell line from embryonic chicken heart tissue. (C) Signup and view all the answers

Which of the following lists the scientists in chronological order based on their contributions mentioned?

Roux, Harrison, Carrel & Burrows, Kohler & Milstein, Steptoe & Edwards (A) Signup and view all the answers

What fundamental shift in biological research is reflected by the transition from Roux's neural fold maintenance to Carrel and Burrows's cell line generation?

A shift from observing tissue fragments to establishing sustained cell populations for study. (C) Signup and view all the answers

In what way did the work of Steptoe and Edwards build upon earlier tissue culture techniques?

They utilized in vitro techniques to enable fertilization and early embryonic development. (B) Signup and view all the answers

Which of the following discoveries had the least direct impact on advancing vaccine development?

The maintenance of neural folds from early chick embryos by Wilhelm Roux. (D) Signup and view all the answers

Considering the progression of tissue culture techniques, what conceptual link connects Harrison's work with frog embryos to Steptoe and Edwards's in vitro fertilization?

Both involved the manipulation of embryonic tissues outside their natural environment to facilitate development. (D) Signup and view all the answers

What is the primary role of the extracellular matrix (ECM) in tissue culture?

To support cell attachment and promote normal cell functioning. (C) Signup and view all the answers

Why is trypsin used in the context of tissue culture?

To break down the extracellular matrix and separate cells. (B) Signup and view all the answers

Which characteristic distinguishes most cell types (excluding blood cells) in tissue culture?

Their need to attach to an extracellular matrix. (B) Signup and view all the answers

What is the significance of coating tissue culture vessels with components of the extracellular matrix (ECM)?

It promotes cell adhesion, growth, and normal functioning of adherent cells. (D) Signup and view all the answers

Which of the following best describes the primary advantage of using tissue culture in neuroscience research?

It allows for observation of cellular behavior in a simplified and controlled environment. (C) Signup and view all the answers

How does the process of 'passaging' cells relate to enzymatic dissociation in tissue culture?

Passaging involves enzymatic dissociation to transfer cells to new vessels. (D) Signup and view all the answers

Why is understanding the historical context of tissue culture important for modern neuroscientists?

It provides insights into previous experimental failures, refining current methodologies. (C) Signup and view all the answers

Why might researchers use purified components of the ECM instead of unpurified components in tissue culture?

Purified components offer better control over the culture environment. (D) Signup and view all the answers

Which aspect of a tissue culture environment is MOST critical for maintaining the viability of neural cells in vitro?

Strict control over temperature, pH, and nutrient supply. (D) Signup and view all the answers

Which of the following is a significant challenge when extrapolating results from neural tissue culture studies to in vivo conditions?

The absence of systemic influences and the simplified environment in culture. (C) Signup and view all the answers

What role do cell adhesion molecules play in tissue culture?

They facilitate the attachment of cells to the ECM. (D) Signup and view all the answers

Consider a scenario where cells in a tissue culture are failing to attach to the culture vessel. Which modification would be most effective at improving cell adhesion?

Coating the culture vessel with purified ECM components. (D) Signup and view all the answers

Consider a researcher observing unexpected cell death in a neural tissue culture. Which initial step would be MOST crucial in troubleshooting the issue?

Thoroughly assessing the culture media, temperature, and other environmental conditions. (C) Signup and view all the answers

Which of the following experimental designs would MOST effectively utilize tissue culture to study the effects of a neurotoxin on neuronal function?

Observing changes in neuronal morphology and activity after exposure to varying concentrations of the neurotoxin. (D) Signup and view all the answers

A researcher observes that cells in a culture are clumping together excessively. Which of the following could be a potential solution to promote a more even cell distribution?

Treating the cells with EDTA, a calcium chelator, before passaging. (C) Signup and view all the answers

A researcher aims to model a neurodegenerative disease in vitro. What key consideration should guide their choice of cell type and culture conditions?

Selecting cells and conditions that closely mimic the affected neurons and the disease's pathological environment. (B) Signup and view all the answers

In a long-term tissue culture experiment, what potential problem does passaging address, and what is a crucial consideration during this process?

Passaging mitigates overcrowding and nutrient depletion; minimizing cellular stress during dissociation. (B) Signup and view all the answers

In a long-term neural tissue culture experiment, a researcher observes a gradual decline in neuronal signaling efficiency. Which factor is LEAST likely to contribute to this decline?

Increased activity, leading to enhanced synaptic connections. (B) Signup and view all the answers

When designing a tissue culture experiment to assess the impact of a novel drug on synapse formation, what control condition is MOST appropriate?

A culture treated with the drug's solvent at the same concentration. (D) Signup and view all the answers

A researcher is using a 3D neural tissue culture to study the migration patterns of newly differentiated neurons. What advantage does this model offer compared to traditional 2D cultures for this specific application?

3D cultures more accurately mimic the complex extracellular matrix and cell-cell interactions present in vivo. (D) Signup and view all the answers

What key advancement did the use of trypsin provide in the context of cell culture, as highlighted by Rous & Jones (1916)?

It allowed for the repetitive subculture (passaging) of substrate-attached cells, which was crucial for expanding cell populations. (D) Signup and view all the answers

Which of the following describes a limitation of using trypsin for cell passaging that has led to the adoption of alternative methods?

Trypsin can alter cell surface proteins and receptors, potentially affecting cell behavior and experimental outcomes. (C) Signup and view all the answers

In the context of cell culture, what is the primary role of the extracellular matrix (ECM)?

To provide structural support and signaling cues that influence cell adhesion, migration, and differentiation. (A) Signup and view all the answers

What is a key advantage of using EDTA solutions as an alternative to trypsin for cell dissociation?

EDTA is a non-enzymatic method that minimizes the risk of damaging cell surface proteins and receptors. (C) Signup and view all the answers

Consider a scenario where cells in culture are failing to adhere properly to the substrate despite appropriate coating. Which of the following factors is LEAST likely to be the primary cause?

Contamination of the cell culture with mycoplasma, which competes with cells for binding sites on the substrate. (D) Signup and view all the answers

In designing a long-term cell culture experiment, what strategy would be MOST effective in minimizing phenotypic drift caused by passaging?

Reduce the number of passages by using lower seeding densities and larger culture vessels to minimize the frequency of enzymatic dissociation. (C) Signup and view all the answers

A researcher observes that cells are detaching from the culture flask prematurely, even though trypsin has not been added. Which of the following is the MOST likely explanation for this observation?

The culture medium lacks sufficient adhesion factors, such as fibronectin or vitronectin. (A) Signup and view all the answers

If a lab is switching from trypsin to Accutase for cell passaging, what is the MOST important consideration for adapting their existing protocols?

Accutase's enzymatic activity might require a longer incubation period to achieve the same level of cell detachment as trypsin. (D) Signup and view all the answers

Which of the following scenarios presents the MOST significant ethical challenge in the application of induced pluripotent stem cells (iPSCs) derived from tissue culture?

Balancing the potential for therapeutic benefits with concerns about the source and procurement of somatic cells for reprogramming. (B) Signup and view all the answers

A researcher observes that cells in their tissue culture are exhibiting decreased proliferation rates, abnormal morphology, and increased cell death following an induced stress. Which of the following interventions would be the MOST comprehensive approach to address these issues and restore optimal culture conditions?

Analyzing the culture medium for nutrient depletion or accumulation of toxic metabolites, coupled with a partial or complete medium change. (A) Signup and view all the answers

In the context of tissue culture, which strategy BEST addresses the challenge of maintaining cellular differentiation in vitro over long periods?

Providing a three-dimensional culture environment that mimics the in vivo tissue architecture. (B) Signup and view all the answers

A researcher aims to establish a long-term culture of primary hepatocytes to study drug metabolism. What is the MOST critical factor to consider for successful maintenance of hepatocyte-specific functions in this in vitro system?

Providing a suitable extracellular matrix and co-culture with non-parenchymal cells to mimic the liver microenvironment. (D) Signup and view all the answers

Critically evaluate the following statement: 'The successful application of tissue culture techniques in regenerative medicine relies solely on the efficient proliferation of cells in vitro.'

The statement is inaccurate because cellular differentiation, maintenance of function, and appropriate tissue organization are also critical for successful regenerative medicine. (D) Signup and view all the answers

What inherent design feature of the Carrel flask most directly mitigates the risk of contamination during brief periods of being open to the environment?

The angled neck, which reduces the settling of airborne particles into the flask. (C) Signup and view all the answers

How did the introduction of antibiotics such as penicillin and streptomycin impact the field of tissue culture?

They diminished the problem of microbial contamination of cultures, enhancing the reliability and longevity of experiments. (B) Signup and view all the answers

In contemporary tissue culture practices, what is the primary advantage of utilizing single-use, sterile plastic vessels compared to reusable glass alternatives?

Reduction in the risk of microbial and cross-contamination. (D) Signup and view all the answers

A researcher is designing an experiment requiring long-term cell culture with minimal disturbance. Which type of vessel would be most appropriate, considering the need for both nutrient availability and contamination prevention?

A large flask (175 cm²) with a vented lid, promoting gas exchange and nutrient availability while reducing contamination risks. (A) Signup and view all the answers

A lab technician discovers fungal contamination in a supposedly sterile culture flask. Considering best practices for preventing future occurrences, what procedural change would most effectively address this issue?

Implementing stricter sterilization protocols for all equipment and consumables, including enhanced air filtration. (C) Signup and view all the answers

A researcher aims to establish a high-throughput screening assay for drug candidates affecting neuronal differentiation. Considering the need for numerous parallel experiments, which vessel format is most suitable?

A plate with 384 wells, enabling a large number of conditions to be tested in parallel with minimal reagent usage. (B) Signup and view all the answers

What is the most critical reason for using vented lids on tissue culture flasks?

To enable proper gas exchange, maintaining optimal levels of $CO_2$ and $O_2$ for cell viability while preventing contaminant entry. (A) Signup and view all the answers

In the context of tissue culture, what is the most significant implication of cross-contamination between different cell lines?

Compromised experimental integrity and unreliable research findings due to the presence of unintended cell types. (D) Signup and view all the answers

A research team is investigating the effects of a novel neurotoxin on neuronal cell cultures. Considering the evolution of tissue culture techniques, what control measure reflects an advancement beyond the methods available in Baker's time (1923)?

Implementing sterile, single-use plastic vessels to prevent cross-contamination. (C) Signup and view all the answers

A cell culture lab is experiencing recurring issues with mycoplasma contamination, despite following standard sterile procedures. Which advanced strategy represents the MOST effective approach to address this persistent problem?

Implementing routine PCR-based screening of all cell lines, combined with immediate quarantine and treatment or disposal of infected cultures. (C) Signup and view all the answers

A researcher aims to create a three-dimensional (3D) cell culture model to better mimic the in vivo environment of a brain tumor. Which methodological consideration is MOST critical for ensuring the model's physiological relevance?

Optimizing the composition of the extracellular matrix (ECM) scaffold to promote appropriate cell-cell and cell-matrix interactions. (A) Signup and view all the answers

In the development of advanced bioreactors for large-scale cell culture, what engineering challenge presents the MOST significant hurdle to overcome?

Ensuring uniform nutrient and oxygen supply to all cells while minimizing shear stress. (D) Signup and view all the answers

A researcher is investigating the long-term effects of a novel drug on neural stem cell differentiation. To ensure the reliability and reproducibility of the study, which practice is MOST crucial?

Implementing rigorous quality control measures to monitor cell viability, phenotype, and genetic stability over time. (A) Signup and view all the answers

Considering the evolution of tissue culture techniques, what ethical consideration has become increasingly prominent with the advent of induced pluripotent stem cells (iPSCs) and their potential for generating diverse cell types?

The importance of obtaining informed consent from tissue donors and addressing potential concerns about commercialization of human biological materials. (C) Signup and view all the answers

A lab is transitioning from traditional 2D cell culture to a 3D bioprinting approach for creating tissue constructs. What factor presents the MOST significant challenge in achieving functional and physiologically relevant 3D tissues?

The difficulty of precisely controlling cell positioning and architecture to mimic native tissue organization. (C) Signup and view all the answers

Flashcards

Tissue Culture

The process of growing cells or tissues in a controlled environment outside of their natural context.

Challenges in Tissue Culture

Maintaining a sterile environment, providing the correct nutrients, regulating temperature and gas exchange, and preventing contamination.

Applications of Tissue Culture

Includes research, drug discovery, toxicity testing, and regenerative medicine.

IPCs

Induced Pluripotent Stem Cells. Cells derived from adult tissues that have been reprogrammed back to an embryonic stem cell-like state.