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Enzymatic Isolated Chondrons from Rabbit Knee Articular Cartilage in Virto

Author: SunZhenWei
Tutor: WeiXiaoChun
School: Shanxi Medical
Course: Orthopaedic Surgery
Keywords: Rabbit knee Chondrocytes Cartilage Unit Enzymatic
CLC: R681.3
Type: Master's thesis
Year: 2010
Downloads: 18
Quote: 0
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Abstract


Objective To investigate the rabbit articular cartilage units (Chondron) in vitro enzymatic digestion , separation feasibility and experimental methods . Method two 8 -month-old New Zealand rabbits were randomly divided into two groups, each of four . Sectional view taken under sterile conditions, double knee full thickness cartilage, a group with conventional 0.4% Pronase enzyme and 0.025% Ⅱ collagenase digestion followed into chondrocytes ; another group with 0.3% dispase enzymes and 0.2% Ⅱ collagenase joint 3h stirring cartilage digestion unit . Using the inverted microscope , cells seeded HE and immunofluorescence staining of collagen type Ⅵ and micro- system test analyzer morphological analysis of cartilage cells and units , Annexin-V/PI cartilage cells by flow cytometry and units of early apoptosis rate . Results inverted microscope unit mainly composed of cartilage , 2, 3 or 4 beaded morphological structure, translucent less degree ; while chondrocytes showed a single translucent rounded cell morphology . HE staining chondrocytes climbing film morphology, cytoplasmic leavening translucent ; cartilage units without significant climbing film , there is a layer of lightly stained cells surrounding the cell cycle matrix components . And matrix components collagen type Ⅵ immunofluorescence positive expression , clear boundaries , showing that a large number of parcels 2, 3 or 4 by the unit cell morphology . Unit smears were divided into cartilage descending nucleus, cytoplasm and cell cycle matrix three-tier structure , each layer gradient height of about 0.4μm. Acute digestive chondrocytes and unit rates in living cells , early and late apoptotic rate was no significant difference . Conclusion 0.3% dispase enzymes and 0.2% Ⅱ collagenase digestion 3h joint agitation rabbit articular cartilage can be successfully acquired units , lay the foundation for further studies in vitro .

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CLC: > Medicine, health > Surgery > Orthopaedic Surgery ( movement system diseases,orthopedic surgery ) > Bone diseases > Cartilage disease
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