Mammalian Cell Culture Overview

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Questions and Answers

Quale test es essential pro assecurar que le medium non es compromittite post filtration?

  • Verificar si le membrana del filtro es intacte (correct)
  • Evaluar le sapor del medium
  • Assicurarse que le pH del medium es correct
  • Testar si le medium es colorate

Que significa 'Plating efficiency' in le contexto de cultura cellulare?

  • Le proportion de cellul que cresce in colonias de celles in 100 celles inoculate (correct)
  • Le velocitate de croissance del celles durante le incubation
  • Le numero de fois que le celles es transferite a un novo medium
  • Le quantitate de medium usate pro cultiva cellulapor

Quale de le sequente es un test de sterilisazione pro le medium?

  • Transferimento de celles a un novo receptacle
  • Observation del color del medium
  • Misura del pH del medium
  • Incubation de aliquotas de medium in varios broths (correct)

Que es le metodo pro comparar le characteristics de cultura cellular inter le novos e vetustos lots de medium?

<p>Compare le curvas de croissance (B)</p> Signup and view all the answers

Quando un medium es preparate in le laboratorio, que tipo de test debe esser realizate ante le uso?

<p>Testo de promtion de croissance e sterilitá (B)</p> Signup and view all the answers

Quande se discute le media de cultura cellular mammalian?

<p>In le octave et novem septiman (D)</p> Signup and view all the answers

Qual de iste non es un metodo de sterilisation pro media de cultura cellular?

<p>Sterilisation per radiomembrana (C)</p> Signup and view all the answers

Qual es un control de qualitate que debe esser facte ante le uso de media?

<p>Controllo del pH (A)</p> Signup and view all the answers

Quande se face un recap de lecturas ante le examenes?

<p>In le undecime et duodecime septiman (B)</p> Signup and view all the answers

Qual lection recuntrara le biologia del cellulas de cultura?

<p>Leccion 8 (C)</p> Signup and view all the answers

Qual es un tema discutite in le lection 15?

<p>Response immunitaria innate (B)</p> Signup and view all the answers

Qual de iste non es un parte del programma de lection?

<p>Culture de cellulas vegetales (A)</p> Signup and view all the answers

Qual es le methodo de sterilisation que non es necessario si le media es preparate in un ambiente aseptic?

<p>Filtration per membrana (C)</p> Signup and view all the answers

Qual broth es usate pro isolar organisimos aerobic e facultative?

<p>TSB (D)</p> Signup and view all the answers

Quo es le principal causa pro sterilisar le media pro cultura cellular?

<p>Proteger le media ab contamination (B)</p> Signup and view all the answers

Quo es un signale de contamination in un sterility test?

<p>Cloudiness (C)</p> Signup and view all the answers

Qual es le temperature correct pro incubar TSB media durante le test de sterility?

<p>37 C (D)</p> Signup and view all the answers

Quale de le sequente techniques es crucial pro mantener le sterilidade del media?

<p>Technique aseptica (C)</p> Signup and view all the answers

Pro quantos dies deberea le SDB media essere incubate in le test de sterility?

<p>5 dies (D)</p> Signup and view all the answers

Quale es le methodo preferite pro sterilisar media pro cultura cellular?

<p>Filtration (C)</p> Signup and view all the answers

Qual broth es usate pro isolar organisimos anaerobico?

<p>TGB (A)</p> Signup and view all the answers

Quale es le pore size recomendate pro le filtrage steril del media?

<p>0.2µm (D)</p> Signup and view all the answers

Qual es le prime passo ante sterilisation de media?

<p>Dissolver in aqua purificate (D)</p> Signup and view all the answers

Quale de le sequente materiales debe esser inert pro un filter sterilizante?

<p>Inoxidable (D)</p> Signup and view all the answers

Quale effetto debe esser confirmate per le studi de crescimento post-filtrage?

<p>Nullo effetto super le proprietates de promovimento del crescimento (B)</p> Signup and view all the answers

Qual es le procedura pro verificar le sterility de un media?

<p>Incubate e observar pro crescita (C)</p> Signup and view all the answers

Quo debe ocurrer si contamination es observate in un batch?

<p>Discarder le batch (C)</p> Signup and view all the answers

Quale de le sequente es un requirement pro le containers de media?

<p>Debent esser steril (D)</p> Signup and view all the answers

Quo non es un criterium pro le media components?

<p>Costo basse (C)</p> Signup and view all the answers

Qual es le fontanian de aqua considerat le plus pur?

<p>Aqua pro injection (B)</p> Signup and view all the answers

Quale de le sequente non es un methodo de purification de aqua?

<p>Ajustamento de pH (D)</p> Signup and view all the answers

Porque aqua potabile debe subire purificazione in-domicilio ante utiliser?

<p>Illa ha contaminantes organicos (D)</p> Signup and view all the answers

Qual parte del processu de purificazione elimina microorganismos?

<p>Filtration (A)</p> Signup and view all the answers

Qual es un detail importante a documentar durante le preparation de media?

<p>Le materiaes prime usate (C)</p> Signup and view all the answers

Quale de le sequente describe aqua purificate?

<p>Le aqua qui ha subite processu de filtration (C)</p> Signup and view all the answers

Qual es le standard US que determina le criteria pro aqua pro injection?

<p>USP (A)</p> Signup and view all the answers

Qual contaminante es eliminat per 'Activated Charcoal'?

<p>Material organico (D)</p> Signup and view all the answers

Flashcards

Sterilisation del medio de cultura cellulaire

Le processo de remover microorganismos del medio de cultura cellulaire per filtrar lo per un membrana con un diametro de pore minus que le microorganismos.

Controles de qualitate del medio de cultura cellulaire

Un methodo de control de qualitate pro verificar si le medio de cultura cellulaire es libere de microorganismos ante usar lo.

Ambiente aseptic pro media de cultura cellulaire

Un ambiente aseptic es un ambiente libere de microorganismos. Un ambiente aseptic es essential pro le preparation del medio de cultura cellulaire.

Sterilisation del medio de cultura cellulaire preparate con ingredientes sterile

Le medio de cultura cellulaire es preparate con ingredientes sterile in un ambiente aseptic. Sterilisar le medio con filtration de membrana post le preparation non es necessari.

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Sterilisation del medio de cultura cellulaire preparate con ingredientes non sterile

Le medio de cultura cellulaire es preparate con ingredientes que non es sterile. Sterilisar le medio con filtration de membrana post le preparation es necessari.

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Verificate del medio de cultura cellulaire pro qualitate

Le medio de cultura cellulaire debe esser verificate pro assicurar que illo es sterile e de bon qualitate.

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Importantia de ingredientes sterile

Le primo passo in le preparation del medio de cultura cellulaire es asegurar que le ingredientes es sterile. Si le ingredientes non es sterile, le medio es probabilmente contaminate.

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Importantia de un ambiente aseptic

Le ambiente ubi le medio de cultura cellulaire es preparate debe esser aseptic - libere de microorganismos. Si le ambiente non es aseptic, le medio es probabilmente contaminate.

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Sterilitate in medio de cultura cellular mammalian

Le medio de cultura cellular mammalian debe esser libere de contaminantes e agentes biologic que pote causar problemas como bacteria, fungi, virus, e mycoplasma.

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Importantia de sterilitate in medio de cultura cellular mammalian

Le medio de cultura cellular mammalian es un ambiente ric pro le crescimento de microorganismos. Pro evitar isto, le technicas de sterilitate es crucial.

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Technicas de sterilitate in medio de cultura cellular mammalian

Technicas de sterilitate in medio de cultura cellular mammalian include laminar flow, superficies de travalio facilemente lavabile, personnel instruite, reagent e medio con controlo de qualitate, technica aseptic, e apparato e equipamento con manutenentia regular.

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Components del medio de cultura cellular mammalian

Le componentes del medio de cultura cellular mammalian debe esser del plus alte qualitate e puritate, e debe esser immagazinate correctemente pro evitar deterioration.

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Containers pro medio de cultura cellular mammalian

Le containers pro medio de cultura cellular mammalian debe esser sterile. Le plus communmente usate es flacon e containers disposable de plastic sterile.

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Autoclavamento de medio de cultura cellular mammalian

Autoclavamento non es un option pro sterilise le medio de cultura cellular mammalian, porque illo pote destruir componentes e biomoleculas crucial pro le crescimento cellular.

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Sterilisation de medio de cultura cellular mammalian: Filtration

Filtration es le methodo preferite pro sterilise medio de cultura cellular mammalian. Filtros de membrana con pore de 0.2µm o minus es usate pro remover particulas.

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Material de construction pro filtros de sterilisation

Le material de construction pro filtros de sterilisation debe esser inerte pro evitar le addition o removal de componentes al medio. Le filtration non debe afectar le proprietates de crescimento del medio o le expression de proteina.

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Filtro sterile in le medio de cultura cellular

Le filtro sterile es un componente critic in le preparation de medio de cultura cellular. Si le filtro es damnate, le medio pote esser contaminate e non apte pro le cultivo cellular.

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Teste de integritate del filtro sterile

Pro verificar si le filtro sterile functiona correctemente, un teste de integritate es executate post le filtration. Iste teste verifica si le membrana del filtro es intacte, e si non, le medio pote esser compromise.

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Teste de promotion del crescimento cellular

Un medium de cultura cellular que es preparate in le laboratorio debe esser testate pro su capacitate de promover le crescimento cellular ante que esser usate.

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Teste de sterilitate in le medio de cultura cellular

Le teste de sterilitate verifica que le medio de cultura cellular es libere de microorganismos no desirabile. Iste teste involve incubar aliquots de medio in diverse broths pro verificar si le bacteria o le levaduras cresce.

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Efficacia de placar in le medio de cultura cellular

Le efficacia de placar es un mesura del numero de cellulas in un population que pote se attachar e crecer a un superficie. Iste teste compara le crescita cellular in le medio nove con le medio vetere pro verificar le consistentia del medio.

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Purification de Aqua potabile

Le aqua potabile es tractade per filtrassion pro remover contaminantes e bacterias, pro que es necessari pro cultivar cellulas mammalian.

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Aqua pro Injection (WFI)

Un typo de aqua que es tractade pro remover quasi omne contaminantes, e es ideal pro le preparation de medio pro cultivar cellulas mammalian.

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Filtration de Profundo

Un typo de filtration que remove particulas solide del aqua usando un membrana con poros microscopic.

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Trappa Organico

Un typo de filtration que remove particulas organico del aqua, como componentes de material organic.

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Charcoal Activate

Un material porose que absorbe e remove contaminantes organico e alge del aqua.

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Exchange de Aniones

Un typo de filtration que remove aniones del aqua, como cloro e bromino.

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Exchange de Cationes

Un typo de filtration que remove cationes del aqua, como calcio e magnesio.

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Distillation / Filtration de Reverse Osmosis

Un processo que remove le major parte del microrganismos del aqua.

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Test de Sterilitate

Un methodo pro testar le sterilitate de medio de cultura cellular. Implica le inoculation de un medio con un cultura bacterial o fungo, e observar le crescimento in le tempore.

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Trypticase Soy Broth (TSB)

Un broth nutriente specific pro le crescimento de bacterias. Il es usate in testar le sterilitate de medio de cultura cellular.

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Sabouraud Dextrose Broth (SDB)

Un broth specific pro le crescimento de fungos. Il es usate in testar le sterilitate de medio de cultura cellular.

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Nebulositate

Le signo visual de contamination in un medio de cultura cellular. Significa que le medio non es plus sterile.

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Aerobios facultative

Un grupo de organismos que pote crescer in le presentia de oxygeno, ma pote anque crescer in absentia de oxygeno. Exemplos include Escherichia coli e Staphylococcus aureus.

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Microaerophilic

Un grupo de organismos que require un ambiente basse in oxygeno pro crescer. Exemplos include Helicobacter pylori e Campylobacter jejuni.

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Anaerobic

Un gruppo de organismos que solo pote viver in absentia de oxygeno. Exemplos include Clostridium botulinum e Bacteroides fragilis.

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Sterilisation per Calor

Un methodo pro sterilizar le medio de cultura cellular per le uso de calor. Le aqua es bullite e vaporisa, e le calor es transferite al medio, desictente omne organismos viventie.

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Study Notes

Week Schedule

  • Week 1: Lecture Module Introduction, Tuesday, September 10th
  • Week 2: Lecture 1 (Use of Mammalian Cells), Monday, September 16th; Lecture 2 (Cell Culture Lab Layout, Equipment, and Materials), Tuesday, September 17th
  • Week 3: Lecture 3 (Contamination Control), Monday, September 23rd; Lecture 4 (Contamination Control), Tuesday, September 24th
  • Week 4: Lecture 5 (Contamination Control), Monday, September 30th; Recap of Lectures 2, 3, 4, and 5, and sample assessment questions, Tuesday, October 1st
  • Week 5: Lecture 6 (Nutrient Uptake), Monday, October 7th; Lecture 7 (Nutrient Uptake), Tuesday, October 8th
  • Week 6: Lecture 8 (Biology of Culture Cells), Monday, October 14th; Lecture 9 (Cell Culture Media), Tuesday, October 15th
  • Week 7: Lecture 10 (Cell Culture Media), Monday, October 21st; Lab 3 Data Analysis, Tuesday, October 22nd
  • Week 8: Reading Week
  • Week 9: Lecture 10 (Cell Culture Media), Monday, November 4th; Lecture 11 (Cell Culture Media), Tuesday, November 5th; Lecture 12 (Growing Mammalian Cells), Monday, November 11th; Lecture 8, 9, 10, and 11 recap and sample assessment questions, Tuesday, November 12th
  • Week 10: Lecture 13 (Monitoring Growth), Monday, November 18th; Lecture 14 (Cryopreservation), Tuesday, November 19th
  • Week 11: Lecture 15 (Innate Immune Response), Monday, November 25th; Lecture 16 (Adaptive Immune Response and Bioassays), Tuesday, November 26th; Recap of Lectures 12, 13, and 14, and sample assessment questions
  • Week 12: Lecture 15 and 16 recap and sample assessment questions, revision, Monday, December 2nd; Revision, Tuesday, December 3rd

Mammalian Cell Culture Media - Lecture Overview

  • Introduction: Why discuss this topic
  • Main discussion: Mammalian cell culture media, sterilization and quality control
  • Conclusion: Take home message

Mammalian Cell Culture Media - Sterility

  • Cell culture media need to be free of harmful contaminants and adventitious biological agents (bacteria, fungi, viruses, mycoplasma).
  • Sterility relies on laminar flow biological safety cabinets / cleanrooms, suitable work surfaces, trained personnel, quality controlled reagents and media, and meticulous aseptic technique and equipment maintenance.

Mammalian Cell Culture Media - Sterilization Methods

  • Membrane filtration is the preferred method for sterilizing cell culture media.
  • Autoclaving is generally not suitable as it could destroy critical components.
  • Sterile filters are rated for 0.2µm or smaller.
  • Filter materials used must be inert and not add or remove components.

Quality Control of Cell Culture Media

  • Growth Promotion: Evaluate growth characteristics by comparing old and new media batches.
  • Sterility Tests: Standard microbiological tests for bacteria/yeasts, incubating aliquots of media in different broths (e.g., Trypticase Soy Broth), incubating and observing for growth.
  • Sterile Filter: Ensure filter membrane integrity is tested to confirm intact membrane after filtration.

Water Purification for Cell Culture Media

  • Components of cell culture media are dissolved in purified water.
  • Potable water is subjected to further in-house purification prior to use.
  • Prepared media are sterilized by filtration and tested for sterility
  • Preparation details should be recorded, and an expiry date assigned to the batch.

Water for Injection (WFI)

  • Water for injection (WFI) is the purest form of water for use in medicinal applications.
  • Distinguished from purified water based on the range/quantity of allowable contaminants.

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