Biology of Cell Culture Lectures Schedule

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Questions and Answers

Qual es le primo passo in le resuscitation de cellulas congelate?

Examinar le cellulas microscopicamente.

Labelar le flasks con le nomine del cellula. (correct)

Incubare le cellulas in un ambiente CO2.

Pipettar le contento del ampoule in un tubo sterile.

Qual metodo es recommandate pro le congelation de cellulas pro garantir le viable?

Congelar cellulas in un medium non pre-calentate.

Congelar rapidamente in un freezer.

Congelar cellulas in le fase log con un alta vitalitate. (correct)

Utilisar un liquido nitrogeno pro congelar immediatemente.

Porque es importante usar un cryoprotectant durante le congelation de cellulas?

Pro augmenter le temperature de congelation.

Pro reducir le tempo de congelation.

Pro impedir le formation de cristallos de glace in le cellulas. (correct)

Pro aumentar le densitate de cellulas.

What should be done to thaw ampoules containing frozen cells?

Place le ampoule in un bano de aqua a 37°C. Signup and view all the answers

Qual es le methodo preferite pro le storage a longo terminos de cellulas?

Usar nitrogeno liquido. Signup and view all the answers

Qual es le temperatura optim pro storage de cellulas in liquid nitrogen?

-196°C Signup and view all the answers

Qual es le concentration minim de serum/protein recomendate pro cryopreservation?

<blockquote> 20% </blockquote> Signup and view all the answers

Qual es le effecto de un cooling rate de 1°C/min?

Incremente le viabilitate de cellulas. Signup and view all the answers

Que debe esser le viabilitate del culturas ante le congelation?

<blockquote> 90% </blockquote> Signup and view all the answers

Qual cryoprotectant es frequentemente usate pro cryopreservation?

Dimethyl sulphoxide (DMSO) Signup and view all the answers

Que debe succeder con le cellulas post congelation pro maximizare le viabilitate?

Thaw them quickly. Signup and view all the answers

Quale es un motivo pro cryopreservation de cellulas?

Contaminatio per microorganismos Signup and view all the answers

Quo sustine le storage de cellulas a ultra-low temperatures?

Le temperatures less que -135°C. Signup and view all the answers

Que debe esser le condition de cellulas ante congelation?

Non debe haber contamination. Signup and view all the answers

Que carecteristica es important pro le congelation de cellulas?

Utilisar un cryoprotectant Signup and view all the answers

Qual cryoprotectant es considerat le plus effective?

Dimetil sulfoxide (DMSO) Signup and view all the answers

Qual es le importantia de un regime de monitorage stricte pro un congelator de nitrogen?

Pro proteger le investment in le contents. Signup and view all the answers

Que concentration de DMSO es plus usualmente usate pro cryopreservation?

10% Signup and view all the answers

Quo que debet ocurrer durante le thawing de cellulas?

Thawing rapide Signup and view all the answers

Qual parte del sistema de congelation es responsable pro garantir le nivel de liquid nitrogen?

Sensores de alto e basso nivel. Signup and view all the answers

Quande es il necessitate de preservar un linea de cellulas?

Si le cellulas es senescent Signup and view all the answers

Qual es le procedura appropriata pro liberar cellulas adherente in le processus de congelation?

Utilisar trypsin/EDTA pro liberar les. Signup and view all the answers

Qua es le criterium optimal pro le viabilitate cellular durante le congela?

90% o plus. Signup and view all the answers

Que significa 'high viability' pro le cellulas ante le congelation?

Cellulas con plus quam 90% de sobrevivente Signup and view all the answers

Que es un disadvantage de non cryopreservar un linea de cellulas?

Costes de replacement alto Signup and view all the answers

Quo debe esser inclus in le systema de inventario pro vessels de storage ultra-low temperature?

Etichetas resistent pro cata ampoule. Signup and view all the answers

Quo debe facer post le cellula haber essite re-suspendite in freeze medium?

Congelar a una taxa controlate. Signup and view all the answers

Quo es le primo passus in le procedura de cryopreservation?

Evaluar le culturas pro contaminazione. Signup and view all the answers

Qual es le methodo pro far un conteo de cellulas durante le processus de congela?

Utilisar un microscopio inverted. Signup and view all the answers

Quando se teneva le introduction del modulo?

Martes 10 de Septembre Signup and view all the answers

Qual es le tema discusse in le lectures del septima semana?

Cryopreservation de cellulas Signup and view all the answers

Qual lecture se discute le 'Cell culture media'?

Lecture 10 Signup and view all the answers

Qual es le data de le lecture sobre 'Nutrient uptake'?

Martes 8 de Octobre Signup and view all the answers

Qual parte del corso es dedicate a le immunitate innata?

Lecture 15 Signup and view all the answers

Quando es le revision programmata?

Lunes 2 de Decembre Signup and view all the answers

Qual topic se discute in le lecture 13?

Monitoring growth Signup and view all the answers

Qual aspect del cryopreservation es discutite in le conclusion?

Long term storage Signup and view all the answers

Study Notes

Weekly Schedule for Lectures and Labs

Week 1 (Tue 10th Sept): Lecture Module Introduction
Week 2 (Mon 16th Sept): Lecture 1 - Use of mammalian cells; (Tue 17th Sept): Lecture 2 - Cell Culture Lab layout, equipment, and materials
Week 3 (Mon 23rd Sept): Lecture 3 - Contamination control; (Tue 24th Sept): Lecture 4 - Contamination control
Week 4 (Mon 30th Sept): Lecture 5 - Contamination control; (Tue 1st Oct): Lectures 2, 3, 4, and 5 recap and sample assessment questions
Week 5 (Mon 7th Oct): Lecture 6 - Nutrient uptake; (Tue 8th Oct): Lecture 7 - Nutrient uptake and sample assessment questions
Week 6 (Mon 14th Oct): Lecture 8 - Biology of Culture Cells; (Tue 15th Oct): Lecture 9 - Cell culture media
Week 7 (Mon 21st Oct): Lecture 10 - Cell culture media (postponed); (Tue 22nd Oct): Lab 3 Data Analysis; Reading Week
Week 8 (Mon 4th Nov): Lecture 10 - Cell culture media; (Tue 5th Nov): Lecture 11 - Cell Culture Media
Week 9 (Mon 11th Nov): Lecture 12 - Growing mammalian cells; (Tue 12th Nov): Lectures 8, 9, 10, and 11 - Recap and sample assessment questions
Week 10 (Mon 18th Nov): Lecture 13 - Monitoring growth; (Tue 19th Nov): Lecture 14 - Cryopreservation and Lectures 12, 13, and 14 - Recap and sample assessment questions
Week 11 (Mon 25th Nov): Lecture 15 - Innate Immune response; (Tue 26th Nov): Lecture 16 - Adaptive immune response and Bioassays
Week 12 (Mon 2nd Dec): Lectures 15 and 16 - Recap and sample assessment questions; (Tue 3rd Dec): Revision

Cryopreservation and Storage of Cells

Lecture Overview: Introduction on the topic, the main discussion on cryopreservation for long-term storage of mammalian cells, and a conclusion summarizing the key takeaway points.
Introduction: Discusses why cryopreservation is necessary, focusing on the value of cell lines as products (e.g., drugs, food products, beverages) and crucial raw materials. The need to protect and ensure a consistent supply for an indefinite period is highlighted.
Why Cryopreserve?: The importance of preserving cell lines due to the potential difficulty, cost, and time associated with replacement. This includes factors such as contamination (by microorganisms or other cell lines), misidentification, equipment issues (incubator failure), freeing up resources, a need for distribution, and cell senescence.
Cryopreservation Process: This process, involving cooling to sub-zero temperatures, aims to minimize intracellular ice crystal formation and reduce damage from high-concentration solutes formed during intracellular water freezing. The process is achieved by freezing cells in the log phase with high (>90%) viability, using controlled slow freezing to allow water to leave the cells, using cryoprotectants, and rapid thawing.
Freezing Medium: The use of cryoprotectants like glycerol or dimethyl sulfoxide (DMSO). DMSO is considered more effective due to its better cell penetration. For monolayer cultures, cryoprotectants are typically removed after the first medium change. Freezing media often include higher serum concentrations.
Successful Cryopreservation and Resuscitation: Emphasizing the need for healthy cultures with high viability (>90%), no contamination, and log phase cells. Importance of slow freezing, rapid thawing and using appropriate levels of serum/protein (e.g. >20%) in the freezing medium are also discussed.
Cooling Rate: The optimal cooling rate (1°C/minute) is crucial for minimizing cellular damage during freezing. The slow cooling method encourages extracellular migration. Use of specialized equipment for freezing (Mr Frosty, CoolCell), initial cooling at -70°C or -80°C, and specific storage at -196°C.
Ultra-low Temperature Storage: The technique of maintaining cells in a suspended state for extended periods at very low temperatures (-135°C or lower), achievable with specialized electric freezers or liquid nitrogen.
Liquid Nitrogen Storage: Liquid nitrogen is the most common method, offering long-term storage. If liquid nitrogen isn't available, storage in a conventional freezer at the lowest possible temperature (at -140°C or -150°C) is recommended.
Liquid Nitrogen Freezer Operation and Maintenance: Care and attention are needed. Importance of a strict monitoring regime and electronic liquid-level alarms are emphasized. Piped liquid nitrogen is used with high and low-level sensors.
Inventory Control: Importance of accurate record-keeping, individually labeling ampoules, recording locations using a spreadsheet, and using controls to ensure that ampoule addition and removal are tracked and recorded.
Cryopreservation Procedure (Harvest): Detailed steps for harvesting cells for cryopreservation, including viewing cultures for contamination, releasing adherent cells, cell counting, centrifugation, re-suspending in cryoprotectant, and cryopreservation.
Thawing Cells: Detailed steps for thawing cells, including preparation of flasks, removing ampoules, wiping the ampoule, transferring the cells to a sterile tube, and adding medium for resuscitation, cell counting, and transfer of the cells to appropriate culture flasks.
Conclusion: A summary of the cryopreservation process, emphasizing the necessity for viability, and the importance of appropriate and proper storage for preservation in long-term storage options (either in electric freezers or, preferably, in liquid nitrogen).

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Description

Este quiz examina le schedula de lectiones e laboratorios pro le biologica de cellulas cultura. Include temas como le uso de cellulas mammalian, control de contamination, e uptake de nutrientes. Prepare te per revisar le notas de cada lection.