TÁMOP-4.1.1.C-13/1/KONV-2014-0001 projekt Biological Research Centre Address: H-6726 Szeged, Temesvári krt. 62. Mail: H-6701 Szeged, POB 521. www.brc.hu „Az élettudományi-klinikai felsőoktatás gyakorlatorientált és hallgatóbarát korszerűsítése a vidéki képzőhelyek nemzetközi versenyképességének erősítésére” program keretében finanszírozott ELŐADÁS KIVONAT CLASSROOM LECTURE HANDOUT financed by the program „Practice-oriented, student-friendly modernization of the biomedical education for strengthening the international competitiveness of the rural Hungarian universities” Dátum / Date: 2017. FEBRUÁR 1. / FEBRUARY 1, 2017 Helyszín / Place: MTA SZBK BIOFIZIKAI INTÉZET, TANÁCSTEREM / LECTURE ROOM, INST. OF BIOPHYSICS, BIOLOGICAL RESEARCH CENTRE SZEGED, TEMESVÁRI KRT. 62. Az előadás címe / Title of the presentation: CO-CULTURE MODELS TO STUDY MAMMALIAN CELL INTERACTIONS Előadó / Speaker: MÁRIA A. DELI
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TÁMOP-4.1.1.C-13/1/KONV-2014-0001 projekt
Biological Research Centre Address: H-6726 Szeged, Temesvári krt. 62. Mail: H-6701 Szeged, POB 521. www.brc.hu
„Az élettudományi-klinikai felsőoktatás gyakorlatorientált és hallgatóbarát korszerűsítése a vidéki képzőhelyek nemzetközi versenyképességének erősítésére”
program keretében finanszírozott
ELŐADÁS KIVONAT
CLASSROOM LECTURE HANDOUT
financed by the program
„Practice-oriented, student-friendly modernization of the biomedical education for strengthening the international competitiveness of the rural Hungarian universities”
Dátum / Date:
2017. FEBRUÁR 1. / FEBRUARY 1, 2017
Helyszín / Place:
MTA SZBK BIOFIZIKAI INTÉZET, TANÁCSTEREM / LECTURE ROOM, INST. OF BIOPHYSICS, BIOLOGICAL RESEARCH CENTRE
SZEGED, TEMESVÁRI KRT. 62.
Az előadás címe / Title of the presentation:
CO-CULTURE MODELS TO STUDY MAMMALIAN CELL INTERACTIONS
Előadó / Speaker:
MÁRIA A. DELI
M.A. Deli February 1, 2017
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CO‐CULTURE MODELS TO STUDYMAMMALIAN CELL INTERACTIONS
Maria A. Deli
Institute of Biophysics
February 1, 2017
„Practice-oriented, student-friendly modernization of the biomedical education for strengthening the international competitiveness of the rural Hungarian universities”TÁMOP-4.1.1.C-13/1/KONV-2014-0001
• Introduction
• Cell culture facility
• Equipments
• Cell culture ware
• Sterilization
• Culture media
• Primary cultures
• Cell lines, differentiation
• Subculture (passage)
• Freezing, storing and thawing cells
• Contamination
INTRODUCTION TO MAMMALIAN TISSUE AND CELL CULTURE
BRIEF HISTORY OF TISSUE CULTURE
• 1907 Ross Harrison frog embryonic tissue
• 1912 Alexis Carrel chick embryo heart
• 1948 Katherine Sanford single cells
• 1955 Harry Eagle culture medium
• 1960s cell lines
• 1979‐1981 different culture media
• Recombinant protein expression and hybridoma technique
• Large scale culturing, industrial production
M.A. Deli February 1, 2017
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CELL CULTURE FACILITY
Floor plan for an ideal high‐use tissue culture facility
CELL CULTURE FACILITY
EQUIPMENTS FOR CELL CULTURECO2 INCUBATOR
Temperature 26‐39 ºC 37ºCGas concentration CO2 concentration 0‐10 % 5 %
O2 concentration 18‐1 %Humidity – water tray
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EQUIPMENTS FOR CELL CULTUREPHASE CONTRAST MICROSCOPE
In‐phase out‐phase image of a cell culture
Phase plate and light annulus alignment
Position of objectives inverted
EQUIPMENTS FOR CELL CULTURELAMINAR FLOW (BIOHAZARD) CABINET
Biohazard cabinet: vertical air flow, HEPA filter, built‐in UV lampprotection of cells and researchers
EQUIPMENTS FOR CELL CULTURE
Desktop refrigerated centrifugeVacuum pumpBottles: used medium
humidity trap
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Cabinet for sterilized glassware and culture dishes
Cells taken from a tissue of a living organism and placed in culture (P0)After the first subculture: secondary culture (P1)After continued passages: cell line
Examples of primary cultures• White blood cells isolation from blood – lymphocytes, monocytes
• Macrophage peritoneal lavage
• Endothelial cells aorta, human coronary vessels, umbilical cord vein,
Thawingworking quickly warming vials at 37 ºC (water bath)washing the cells – removal of cryoprotectors
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MICROBIAL CONTAMINATION OF CULTURES
Sporulating mold colony (60 X)
Bacterial contamination (SEM, 300 X)
Edge of mold colony (300 X, SEM)
Budding yeast (300 X, SEM)
Elimination of yeasts: nystatin, amphotericin‐BElimination of molds: nystatin, amphotericin‐BElimination of bacteria: penicillin, streptomycin, gentamycin, etc.