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Cellculture

1,952 bytes removed, 09:19, 13 December 2006
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<p>세포배양 (cellculture)&nbsp;</p>
<p align="center">&nbsp;배 &nbsp;&nbsp;양</p>
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<td bordercolor="#ffffcc" width="793971" bgcolor="#ffffcc" colspan="2" height="96">&nbsp;&nbsp;생체 조직을 생체 밖의 환경에서 어느 일정 기간 생육시키는 기술이다. 생명현상 해명을 위하여 세포, 조직을 독립된 생명체로 인식하고 분자 생물학 등 연구재료로 이용하기 위해 만들었다.&nbsp; 사용구조에 따라 [[기관배양]](organ culture), [[조직배양]](tissue culture), [[세포배양]](cell culture)으로 나눌수있고 세포종류에 따라 [[초대배양]](primary culture), [[세포주 배양]](cell line culture)로 나눌수있다.&nbsp;</td>
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<p align="center">&nbsp;배양의 분류</p>
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<p align="center">초대배양</p>
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<p>&nbsp; 정상조직이나 암조직으로 부터 바로 분리된 세포의 배양으로서 여러 세포종이 혼합된다. 원래의 세포기능을 유지한다는것이 장점이고 세포의 균일성, 재현성을 확신할 수 없다는것이 단점이다.</p>
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<p align="center">세포주 배양</p>
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<p>&nbsp; 무한 증식을 할 수 있는 세포로 현재는 단일 세포라는 개념이다.</p>
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<p align="center">&nbsp;배양시설</p>
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<td width="793" colspan="2" height="45"> <table style="WIDTH: 824px; HEIGHT: 533px" height="533" cellspacing="1" cellpadding="1" width="824" summary="" border="0"> <tbody> <tr> <td bordercolor="#66ffff" width="223207" bgcolor="#66ffff" height="277"> <p align="center">배양설비</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="27">&nbsp;사용용도</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="102"> <p align="center">탄산가스 [[배양기]](CO<sub>2</sub> incubator)</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="102">&nbsp; pH조적은 배지의 중탄산 나트륨과 CO<sub>2</sub>가스의 평형에 의해 이루어진다.<br /> &nbsp; 5% CO<sub>2</sub> - 95% air 유지<br /> &nbsp; 재킷형 CO<sub>2 </sub>incubator 주로 사용 - water 재킷(멸균수) : 온도 조절<br /> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; - 가습 vat : 습도 조절</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="51"> <p align="center">배양실과 [[clean&nbsp;bench]]</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="51">&nbsp; 배양실&nbsp;:&nbsp; 무균실<br /> &nbsp; clean bench : 무균조작을 위한 무균상자,&nbsp;자외선 멸균장치 포함</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="88"> <p align="center">[[배양 접시]]</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="88">&nbsp; culture flask - 배양용량이 큰 배양용기, screw cap부착<br /> &nbsp; petri-dish - 조직배양용 dish(접착세포 배양), 일반세균 배양용dish(부유세포 배양)<br /> &nbsp; multi-well plate - 96 well 사용, 소량의 배양액</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="27"> <p align="center">에어컨</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="27">&nbsp; 온도조절</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="31"> <p align="center">[[Inverted microscope]]</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="31">&nbsp; 무염색 세포관찰</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="28"> <p align="center">[[가압 증기 멸균기]](autoclave)</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="28">&nbsp; 멸균장치</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="27"> <p align="center">[[Deep freezer]]</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="27">&nbsp; 동결보존 -80도 유지</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="27"> <p align="center">[[원심분리기]]</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="27">&nbsp; 세포와 배양액의 분리</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="28"> <p align="center">냉동냉장고</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="28">&nbsp; 배지, 시약 등의 보존</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="27"> <p align="center">[[cover slip]]</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="27">&nbsp; 염색용</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="27"> <p align="center">[[auto pipette]]</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="27">&nbsp; 분주</td> </tr> <tr> <td bordercolor="#66ffff" width="223" bgcolor="#66ffff" height="27"> <p align="center">혈구 게시판</p> </td> <td bordercolor="#ffffcc" width="560" bgcolor="#ffffcc" height="27">&nbsp; 세포수 측정</td> </tr> </tbody> </table>
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<p>&nbsp;사용용도</p>
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<p align="center">탄산가스 [[배양기]]<br />
(CO<sub>2</sub> incubator)</p>
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<p>pH조적은 배지의 중탄산 나트륨과 CO<sub>2</sub>가스의 평형에 의해 이루어진다.<br />
&nbsp; 5% CO<sub>2</sub> - 95% air 유지<br />
&nbsp; 재킷형 CO<sub>2 </sub>incubator 주로 사용 - water 재킷(멸균수) : 온도 조절<br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; - 가습 vat : 습도 조절</p>
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<p align="center">배양실과 [[clean&nbsp;bench]]</p>
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<p>&nbsp; 배양실&nbsp;:&nbsp; 무균실<br />
&nbsp; clean bench : 무균조작을 위한 무균상자,&nbsp;자외선 멸균장치 포함</p>
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<p align="center">[[배양 접시]]</p>
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<p>culture flask - 배양용량이 큰 배양용기, screw cap부착<br />
&nbsp; petri-dish - 조직배양용 dish(접착세포 배양), 일반세균 배양용dish(부유세포 배양)<br />
&nbsp; multi-well plate - 96 well 사용, 소량의 배양액</p>
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<p align="center">에어컨</p>
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<p>&nbsp;&nbsp;온도조절</p>
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<p align="center">[[Inverted microscope]]</p>
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<p>&nbsp;&nbsp;무염색 세포관찰</p>
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<p align="center">[[가압 증기 멸균기]](autoclave)</p>
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<p>멸균장치</p>
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<p align="center">[[Deep freezer]]</p>
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<p>&nbsp; 동결보존 -80도 유지</p>
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<p align="center">[[원심분리기]]</p>
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<p>세포와 배양액의 분리</p>
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<p align="center">냉동냉장고</p>
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<p>배지, 시약 등의 보존</p>
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<p align="center">[[cover slip]]</p>
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<td width="762" height="7">염색용</td>
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<p>[[auto pipette]]</p>
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<td width="762" height="7">분주</td>
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<td width="207" height="7">혈구 게시판</td>
<td width="762" height="7">세포수 측정</td>
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<p align="center">&nbsp;계산법</p>
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<p align="center">&nbsp;&nbsp;전세포수 계산<br />
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<td bordercolor="#ffffcc" width="560762" bgcolor="#ffffcc" height="51">&nbsp;&nbsp;culter counter<br />
&nbsp; 혈구 계산판<br />
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<p align="center">생세포수 계산</p>
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<td bordercolor="#ffffcc" width="560762" bgcolor="#ffffcc" height="91">&nbsp; 원리 : 색소 배제 능력&nbsp;&nbsp; tryphan blue염색 - 사세포는 배제 능력이 없고 청색으로 염색한다.<br />
&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 생세포율(%)&nbsp; = (생세포수) / (생세포수 + 사세포수)<br />
&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &nbsp;생세포수(cell / ml) = 전세포수(cells / ml) X 생존율(%) </td>
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