细胞分离培养与保存技巧.ppt

Immunofluorescent localization of microfilaments and microtubules in mouse oocytes with and without DMSO treatment. Arrowheads indicate F-actin caps. Green, microfilaments; red, microtubules; blue, DNA. Bar = 20?μm. Representative images from three replicates are shown. Effect of cryoprotectants on the asymmetric cell division of mouse oocytes. Rate of large polar body extrusion after treatment with various concentrations of DMSO, ethylene glycol, or glycerol. EG, ethylene glycol. 24 25 26 27 28 -1 0 2 5 Time after freeze-thaw (min) Cell radius (?M) P0.0001 leading to an increase of net-permeable surface area of 1,174 ± 69.8 μM2 per cell 细胞分离培养与保存技巧 细胞微环境（cell niches） 细胞的所有功能包括形成、增殖、存活、动员、归巢、分化、功能展现均与微环境有关 细胞微环境（cell niches） 体液 细胞间质 旁分泌 神经 细胞-细胞 相互作用 理化因素 体液 细胞间质 旁分泌 神经 细胞-细胞 相互作用 理化因素 细胞培养 --控制和调节细胞微环境使之适合细胞生存 培养条件：O2、N2、CO2、温度、湿度、培养皿（瓶）、细胞外基质 培养液组份：离子、糖、血清、pH等 周围细胞：单一细胞、混合细胞 避免污染：细菌、真菌等 Components Molecular Weight Concentration (mg/L) mM Amino Acids Glycine 75.0 30.0 0.4 L-Alanyl-Glutamine 217.0 862.0 3.9723501 L-Arginine hydrochloride 211.0 84.0 0L-Cystine 313.0 48.0 0L-Histidine hydrochloride-H2O 210.0 42.0 0.2 L-Isoleucine 131.0 105.0 0.8015267 L-Leucine 131.0 105.0 0.8015267 L-Lysine hydrochloride 183.0 146.0 0.7978142 L-Methionine 149.0 30.0 0L-Phenylalanine 165.0 66.0 0.4 L-Serine 105.0 42.0 0.4 L-Threonine 119.0 95.0 0L-Tryptophan 204.0 16.0 0.078431375 L-Tyrosine 181.0 72.0 0L-Valine 117.0 94.0 0.8034188 低糖DMEM成份 Vitamins Choline chloride 140.0 4.0 0.028571429 D-Calcium pantothenate 477.0 4.0 0.008385744 Folic Acid 441.0 4.0 0.009070295 Niacinamide 122.0 4.0 0.032786883 Pyridoxine hydrochloride 206.0 4.0 0.019417476 Riboflavin 376.0 0.4 0.0010638298 Thiamine hydrochloride 337.0 4.0 0.011869436 i-Inositol 180.0 7.2 0.04 Inorganic Salts Calcium Chloride (CaCl2-2H2O) 147.0 264.0 1.7959183 Ferric Nitrate (Fe(NO3)39H2O) 404.0 0.1 2.4752476E-4 Magnesium Sulfate (MgSO4-7H2O) 246.0 200.0 0.8130081 Potass