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#primary cells

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Animal Tissue Culture 1

Primary cell culture is defined as:

Primary cell culture approach involves direct isolation from intact adult or fetal tissue within hours after sacrifice maintaining in vivo characteristics including diploid karyotype forty six chromosomes for human cells, tissue-specific enzyme expression such as cytochrome P450 3A4 in hepatocytes, albumin secretion, surfactant production in alveolar type II cells, and physiological hormone responsiveness via appropriate receptors, features progressively lost upon immortalization transformation. Protocol begins sterile dissection under laminar flow hood, mincing organ into one millimeter cubes

Ref: Freshney Primary culture Hayflick limit contact inhibition Hippo YAP; Alberts MBoC diploid tissue explant collagenase dispersal.

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Basic Applied Biology

Primary cell culture is defined as:

Primary cell culture defines culture establishment directly from living tissue obtained via biopsy, surgical resection, embryonic dissection or enzymatic disaggregation without prior in vitro passage or immortalization step, preserving in vivo characteristics. Tissue fragments minced to one cubic millimeter pieces, digested with collagenase type I, trypsin-EDTA, dispase or liberase cocktails to release single cells, filtered to remove debris and seeded into tissue culture flasks containing medium supplemented with serum providing growth factors, antibiotics preventing contamination and attachm

Ref: Freshney Culture of Animal Cells 7th ed. Ch 11 Primary culture; Alberts Molecular Biology of Cell Ch cell culture methods.

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