BME 210 Lecture 11 Cell Culture

BME 210 Lecture 11 Cell Culture - 11. Cell culture Ways to...

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11. Cell culture
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Ways to study cells 1. In tissue slices Cells are fixed, studies limited to structure 2. In living tissue Functional studies are possible. Access to cells is limited 3. Isolated cells Much more flexibility but studies are short-term (few hours) 4. Cell cultures Cell Culture - a set of cells maintained outside of the living animal ( in vitro ) for more than ≈24 hours
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Types of cell cultures 1 Primary Cell Culture Prepared from cells just derived from an animal tissue Cells undergo no or only few divisions Cell survive for a relatively short period of time (days – weeks) Examples: neurons, cardiac myocytes Cell Line If cells divide in primary culture, they can be diluted and re-plated in new vessels – this becomes a “cell line” Cell line grows for a number of generations (< ≈60), then they stop dividing ( in vitro cell aging) Examples: fibroblasts, epithelial cells Immortal Cell Line Some cells (e.g., tumor or stem cells) can divide indefinitely. They can make an ”immortal” cell line Some primary cultures or cell lines can be transformed into immortal cell lines by genetic engineering (e.g., transfection with Epstein-Barr virus)
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Types of cultures 2 1. Anchorage-dependent cell cultures • Require a substrate, a solid surface or other cells for attachment and growth • Majority of cells derived from solid tissues (muscle, neuron, fibroblast, epithelial) • Typically, cells exhibit contact inhibition of growth and movement cells usually grow in monolayers 2. Suspension cell cultures • Grow as cell 3D clusters suspended in solution • Mostly hemopoietic cells and stem cells
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Immortal cell line derived from tumor cells of woman He nrietta La cks. One of the most common laboratory cell lines HeLa cell line
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Cell culture environment Cultures are grown in small Petri dishes or larger flasks in an incubator The incubator provides constant temperature, 100% humidity (to avoid evaporation) and controlled atmosphere (typically filtered room air with 1-10% CO 2 ) Two of the most important factors are the growth media and the growth substrate (for the anchorage-dependent cultures)
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Growth media The composition is tailored for each cell type. Typical components: Inorganic salts – provide electrolyte balance similar to blood Sodium bicarbonate (NaHCO 3 ) – provides pH buffer together with CO 2 in the incubator Glucose – source energy Amino acids – for protein synthesis Vitamins – cofactors in biochemical reactions Antibiotics – prevent infections pH indicator (gives red color) Serum
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BME 210 Lecture 11 Cell Culture - 11. Cell culture Ways to...

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