Lecture 1 Introduction

 Lecture 1 Introduction

  • Primary explant of tissue
  • History
    • Harrison 1907
      • Frog embryo nerve fiber outgrowth in vitro (lymph=nutrient/food)
      • Hanging drop technique
    • Carrel 1912
      • Chick CT explants (embryo extract + blood plasma)→immortal
      • grow in Carrel flask under strict aseptic control until 1946
    • Rous & Jones 1916→trypsinization (detach cells with trypsin into suspension) & subculture of explants
    • Earl et al 1943→1st continuous cell line
    • Keilova 1948→antibiotic use in culture
    • Gey at al 1952→HeLa = 1st human continuous cell line
    • Eagle 1955→defined media (no nutrient)
    • Hayflick & Moorhead 1961→finite (limited) lifespan of human cells (50 gen)
    • 1970→laminar flow cabinet
    • 1980→regulation of gene exp
    • 1998→culture of human embryonic stem cells
    • 2000s→Human Genome Project: genomics, proteomics, tissue engineering (regenerative med)
  • Application
    • Basic
      • IC activity→DNA transcription, protein synthesis, energy metabolism, drug metabolism, cell cycle, differentiation, apoptosis
      • IC flux→RNA processing, hormone receptors, metabolite flux, calcium mobilization, signal transduction, membrane trafficking
      • genomics→genetic analysis, transfection, infection, transformation, immortalization, senescence
      • proteomics→gene products, cell phenotype, metabolic pathways
      • c-c interaction→morphogenesis, paracrine control, cell proliferation, kinetics, metabolic cooperation, cell adhesion & motility, matrix interaction, invasion
    • Applied
      • cell products→biotech, bioreactor design, product harvesting, downstream processing
      • immunology→cell surface epitope, hybridomas, cytokines & signaling, inflammation
      • pharmacology→drug action, ligand receptor interactions, drug metabolism & resistance
      • tissue engineering→tissue constructs, matrices & scaffolds, stem cell sources, propagation, differentiation
      • toxicology→infection, cytotoxicity, mutagenesis, carcinogenesis, irritation, inflammation
  • In vitro vs in vivo
    • dissociation of cells from a 3D geometry to their propagation on a 2D substrate
    • represent only 1@2 cell types
    • culture environment lacks systemic components involved in homeostatic regulation
    • energy metabolism in vitro occurring largely by glycolysis
  • Adv
    • control of environment
    • homogeneity
    • characterization of sample
    • economy
    • mechanization
    • reduction of animal use
    • preservation
    • in vitro modeling of in vivo conditions (3D culture)
  • Limitation
    • expertise
    • quantity of effort, cost and materials
    • dedifferentiation
    • instability
  • Types
    • primary explant→tissue fragment + solid (flask)-liquid (media)
    • cell culture
    • organ culture→tissue fragment + air-liquid; 3D
    • organotypic culture→3D culture with differ cells from synthetic organ

Comments

Post a Comment

Popular posts from this blog

KM

7. suruhanjaya, peranan, fungsi, sprm, spr, beza dng kementerian 006, 008 SPR: badan pengendali perjlnan pr samada di peringkat Dewan Rakyat (Parlimen) mahupun Dewan Undangan Negeri DUN dibentuk pd 4/9/1957 di bawah perutkan perkara 113,114 perlembagaan persekutuan peranan: mengendali sesebuah pr  menjlnkan semakan persempadanan kawasan2 pr parlimen & negeri x kurng dr 8 tahun selepas tarikh siapnya urusan kajian & persempadan semula yg terdahulu  jlnkan urusan pendaftaran serta menyemak & kemaskini senarai nama2 daftar pemilih bukan saja mendaftar pemilih yg baru tapi juga mengeluarkan nama pemilih yg telah tinggal dunia dng kerjasama JPN - hanya pemilih2 berdaftar berhak mengundi bertanggungjawab mengendalikan pr umum & rakyat kecil yakni melibatkan pemilihan ahli2 Dewan Rakyat & DUN termasuklah pr kecil jika ada kekosongan  proses: pengeluaran writ - penamaan calon - kempen pr - pengundian - penjumlahan rasmi undian - pegumuman keputusan  p...
Read more

Final Cell Culture

Image
Cell Counting = process of determining cells no 算细胞 importance, method, which method recommended importance of cell counting ensure accurate seeding densities 播种密度 monitors growth rates 成长 evaluates cytotoxicity or proliferation 细胞毒性与增殖 standardize experimental conditions  methods manual counting hemacytometer slide  requires a minimum of 1x10^6 cells/mL use trypan blue for viability test  viable cells not stained  cheap slow prone to error low sensitivity  electronic counting  original resistance-based counter based on the change in current generated when a cell passes through a narrow orifice 孔 image analysis software  scan stained & unstained cells in a special counting chamber & view  continuous monitoring of living cultures  work by counting cells or determine residual available growth area  rapid & low inherent 固有 error - more accurate  needs single cell suspension  flow cytometry  count by light scat...
Read more

HEMA TEST 3

001 1. role of endothelial cells AT - prostacyclin, NO, TFPI, tPA, uPA PT - VWF, PAI, TAFI 2. platelet structure peripheral = glycocalyx sol gel = utubule coil, open canalicular, dense tubular inner organelle BM produce plat 7-10d TPO in liver, kidney, BM lifespan 8-12d 3. primary hemostasis adhesion activation aggregation  4. VWF cleave by ADAM  bind site: collagen, plat, heparin, FVIII synthesis site: endothelial & megaK plat adhesion, protect FVIII 002  1. coagulation cascade/secondary hemostasis extrinsic/initiation TF+VIIa intrinsic/amplification XII=XI=IX common/propagation X to Xa  PT to T = thrombin burst fibrinogen to fibrin 2. natural inhibitor C&S = VIIIa, Va, PAI inhibitor TFPI = extrinsic AT = thrombin, intrinsic & common inhibitor 3. fibrinolysis  plasminogen to plasmin by tPA, uPA ((PAI inhibit tPA, uPA ((a2 antiplasmin & macroglobulin inhibit plasmin fibrin clot to FDP  ((fibrin inhibit by TAFI clear by RES ((thrombin + throm...
Read more