헬름홀츠 협회의 이니셔티브 및 네트워크 기금, 그 밖의 Kroener-는 Fresenius 재단에서 부여하고, 도이치 Forschungsgemeinschaft에서 부여를 통해 이미징 및 치료 환경 대사 질환, (-이 작품은 헬름홀츠 얼라이언스 ICEMED으로 (부분적으로) 투자되었다 SFB 655 : 조직에 세포).

이 작품은 Helsinski의 선언과 ARVO 동물 정책을 준수합니다. 드레스덴 대학의 동물 시설의 지침에 따라 동물 조직 수집에 사용 된 모든 관련 방법은 미행했다. 동물은 취급 및 보관 실험 동물의 사용 및 관리에 대한 독일 연방 지침에 따라, 그리고 연구는 Landesdirektion 드레스덴에 의해 승인되었습니다했다. 적절한 안락사 방법에 대한 교육 기관의 동물 정책 (IACUC 또는 다른 보드)에 문의하십시오. 특정 주식 및 시약의 작업 농도는이 프로토콜에 수반되는 시약 테이블에서 찾을 수 있습니다. 1. 배양 접시 준비 <ol><li> 75 μg의 충분한 볼륨 코트 요리 / ㎖ 폴리-L-오르니 틴 (즉, 2 ㎖ / 잘 6 잘 플레이트의) 하룻밤 2 일전 예약 해부 현재까지 세포 배양 인큐베이터에서. </li><li&gt; 기음 폴리-L-오르니 틴 다음날 5 분 동안 PBS 3 ㎖와 잘 배 각각 세척. </li><li> 최종 세척을 제거한 다음 판에 피브로넥틴 (희석 1:250, PBS에 4 ㎍ / ml)에 추가하고 하룻밤 인큐베이터에서 접시를 놓습니다. </li><li> 해부의 날에 피브로넥틴을 기음과 요리를 PBS로 2 배 씻는다. 여전히 해리 조직 배양을위한 준비가 될 때까지 최종 PBS가 인큐베이터에 씻어 포함 된 번호판을 반환합니다. 이 때, 조직을 도금하는 PBS 사전을 제거합니다. </li><li> 최대 3 주 동안 인큐베이터를 폴리-L은-ornithinein 코팅 스토어 플레이트. 피브로넥틴으로 코팅 된 플레이트는 최대 1 주 동안 저장 될 수있다. 긴급한 경우 피브로넥틴 코팅은 거의 같은 2 시간에서 수행 될 수있다. 피브로넥틴은 변성에 취약하며 솔루션을 선동하거나 요리를 건조하게하지 않는다. </li></ol> 2. 해부 / 도금 <ol><li> 이 프로토콜은 3 성인 쥐 (연령 3 ~ 6 개월)의 사용에 적용됩니다. </li><li> CHIL내가 전에 쥐를 안락사에 얼음 블록을 구획 뇌. </li><li> 얼음에 추가의 bFGF와 N2 매체의 5 ML을 포함하는 15 ML 팔콘 튜브를 놓습니다. </li><li> 교육 기관의 수의 정책에 따라 쥐를 안락사. </li><li> 머리의 중간 선 아래로 절개하여 두개골에서 뇌를 제거합니다. 신중하게 추출 할 수있는 두뇌를 허용 뼈를 벗겨 집게를 사용합니다. </li><li> 얼음 차가운 PBS를 포함하는 10cm 페트리 접시에 두뇌를 놓습니다. 이것은 남아있는 머리카락과 혈액을 제거합니다. (그림 1A) </li><li> 냉장 단면 블록에 두뇌를 놓습니다. (그림 1B) </li><li> 그림 1C에서 그림과 같이 블록에 하나의 새로운 깨끗한 면도날을 삽입합니다. </li><li> 그림 1D에 도시 된 바와 같이 처음에 두 번째 깨끗한 면도날 3mm의 꼬리를 삽입합니다. </li><li> 조심스럽게 그들과 함께 관심있는 부분을 고려하여 블록에서 블레이드를 들어 올립니다. </li><li> 초에게 플로트PBS에 침지시켜 블레이드 떨어져 위치시킵니다. (그림 1E) </li><li> 관심 영역에서 수확 조직 # 5 집게를 사용하여 (이 예제에서 우리는 앞쪽에 접합면 (전방) ACA, 그림 층을 사용)과의 bFGF를 포함하는 1 ㎖ N2 매체를 포함하는 15 ML 원뿔 튜브에 수집합니다. </li><li> 멸균 조건 하에서 프로토콜의 나머지를 계속 층류 세포 배양 후드로 튜브를 이동. </li><li> 기계적 P1000 피펫의 부속 1 ㎖ 피펫 팁을 사용하여 조직을 해리. 상하 피펫 여러 번 (약 20X 초당 1 피펫 사이클의 속도로) 원뿔 튜브 (도 1G 1-3)의 아래에 대 피펫 팁의 개구를 누르면서. 용액이 균질 될 때 분쇄를 중지합니다. </li><li> 이 솔루션은 어떤 큰 nondissociated 조직이 바닥에 정착 할 수 있도록 2 분 동안 앉아 수 있습니다. (그림 1G 4) </li><li> homogen 수집eous 뜨는 및 N2 매체의 8.5 ML 플러스 20 NG / ML bFGF를, 500 NG / ML Dll4, 500 NG / ML ANG2, 200 nMJAK 억제제를 포함하는 원뿔 튜브에 배치합니다. </li><li> 3 희석 세포 솔루션의 ML / 잘 사용하여 6 - 웰 플레이트의 3 우물에 그들을 플레이트. </li><li> 37 ° C에서 5 % CO 2, 5 % O 2에 가습 세포 배양기에 플레이트를 놓습니다. </li></ol> 3. 데일리 케어 <ol><li> 도금 후 24 시간을 억제제의 bFGF, ANG2, Dll4 및 JAK를 포함 N2 매체와 배양 접시에 매체를 교체합니다. </li><li> 의 bFGF, Dll4, ANG2, 다음 날 JAK 억제제의 알약을 추가합니다. </li><li> 미디어의 변화 10-14 일 총 요소 추가이 교류 과정을 계속합니다. </li></ol> 4. 분화 유도 <ol><li> 매체를 포함 억제제의 bFGF, Dll4, ANG2 및 JAK을 철회하고 추가 요소를 포함하지 않는 N2 매체로 교체합니다. </li><li> 격일 매체를 변경합니다. </li><li> 10 일 이후 세포를 수정하고 면역 세포를 수행합니다. </li></ol> 5. 면역 세포 화학 감지 <ol><li> 매체를 대기음 20 분 동안 4 % 파라 포름 알데히드의 2 ㎖의 세포 배양 접시를 고정합니다. </li><li> 5 분마다 PBS의 3 ㎖로 세척 배. </li><li> 대기음 PBS는, 투과 한 후, 20 분 동안 5 % 정상 당나귀 혈청 (NDS) 및 0.1 % 트리톤 X-100을 함유하는 PBS 중 2 ㎖를 첨가하여 세포를 차단. </li><li> 5 % NDS를 포함하는 PBS의 기본 항체 믹스를 준비합니다. TuJ1 (클래스 III β-튜 불린은), GFAP 및 CNPase 항체 분화 마커 트리플 염색을 위해 함께 사용될 수있는 반면 반대로 네 스틴 항체으로 SC를 식별하기 위해 사용될 수있다. </li><li> 차단 솔루션을 기음과 각 웰에 차 항체 혼합물의 1.5 ML를 추가합니다. 90 분 동안 실온에서 품어. </li><li> 기본 항체를 제거하고 PBS의 3 ㎖의 2 배를 씻어 5 분마다 5 % NDS를 포함하는 PBS의 3 ㎖로 1x는. </li><li> Prepar전자 PBS의 보조 항체는 5 % NDS를 포함. </li><li> 최종 세척을 기음과 각 웰에 차 항체 솔루션의 1.5 ML를 추가합니다. 실온에서 40 분 동안 어둠 속에서 배양한다. </li><li> 차 항체 솔루션을 제거하고 새로 제조 DAPI의 얼룩 2 ㎖ (PBS 500 겨 / ㎖)를 추가합니다. 3 분 동안 품어. </li><li> DAPI 솔루션을 대기음 후 5 분마다 PBS의 3 ㎖로 3 회 세척한다. 세포는 이제 형광 현미경으로 시각화 될 수있다. </li></ol>

<ol>
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Neuroinflammation. 6, 39 (2009).</li><li>Androutsellis-Theotokis, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiogenic+factors+stimulate+growth+of+adult+neural+stem+cells.">Angiogenic factors stimulate growth of adult neural stem cells.</a> PLoS One. 5, e9414 (2010).</li><li>Park, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hes3+regulates+cell+number+in+cultures+from+glioblastoma+multiforme+with+stem+cell+characteristics.">Hes3 regulates cell number in cultures from glioblastoma multiforme with stem cell characteristics.</a> Sci. Rep. 3, 1095-1010 (2013).</li><li>Poser, S. W., Shostak, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ch.+5.">Ch. 5.</a> Cancer Stem Cells - The Cutting. , 89-110 (2011).</li><li>Androutsellis-Theotokis, A., Rueger, M. A., Mkhikian, H., Korb, E., McKay, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signaling+pathways+controlling+neural+stem+cells+slow+progressive+brain+disease.">Signaling pathways controlling neural stem cells slow progressive brain disease.</a> Cold Spring Harb. Symp. Quant. Biol. 73, 403-410 (2008).</li></ol>

우리는 공개 아무것도 없어.

성공적으로 분리, 확장, 성인의 뇌에서 신경 줄기 세포의 분화에 중요한 단계의 대부분은 표준 조직 배양 기술에 공통으로 공유됩니다. 명심하는 목적은 성인의 뇌에서 분리 NSCs의 가능한 자신의 생체 내 특성 (그림 2,1)만큼을 유지해야한다는 것입니다. 종종 필요한주의와 적절한 속도 사이에 균형을 공격해야합니다. 두개골에서 뇌의 분리와 관심은 관심 영역의 식별이 훨씬 ?...

한 세기 이상 전에 라몬 Y.를 Cajal에 의해 뇌 cytoarchitecture의 기본 관찰에 의해 규정 신경 생물학의 중심 교리는, 사춘기가 CNS 1에서 발견되는 신경 네트워크의 복잡성을 주어진 후 신경이 어려울 것을 가졌다. 1960 년대에 알트만의 작품, 그리고 나중에 카플란, 3 H-티미 딘이 실제로 뉴런은 성인 뇌의 서로 다른 영역에서 생성되고 있었다 나타내는 성숙한 신경 세포에서 발견 될 수 있다는 것을 입증에도 불구하고, 교리는 2, 3을 유지하는 것을 계속했다. 증거는 가수 두뇌 4에 존재하는 뉴런의 수의 계절적 변화를 설명 Nottebohm의 연구를 탑재하는 것을 계속했다. 그것은 1999 년까지되지 않았을 때 쥐의 연관 학습 과제의 성능뿐만 아니라 Kornack 및 Rakic ​​시연의 관측과 해마 증가 뉴런의 생성에 굴드 등. 출판 일딸랑 딸랑 소리는 덜 엄격한, 더 플라스틱 뇌의 개념이, 5 인식 된 성인 짧은 꼬리 원숭이, 6에 신경을 계속했다. 이 새로이 생성 된 신경 세포의 세포 소스에 대한 검색은 7 벽감으로 불리는 뇌의 영역에있는 줄기 세포 (SC에)의 이산 인구의 발견으로 이어질. subventricular 영역과 해마의 하위 세부 영역은 두 가지 주요 신경성 지역 8,9로 간주됩니다. 이 위치에서 분리 된 세포는 고전적인 배아 또는 태아의 파생으로 SC의 특성,자가 재생과 분화 가능성을 표시합니다. 신경줄 기세포 (NSCs의)의 경우, 이들은 뉴런, 아스트로 사이트, 및 희소 돌기 아교 세포로 분화 될 수있다. 또한, 이러한 SCS는 이러한 중간 필라멘트 단백질 네 스틴 10 태아 NSC 마커에 대한 긍정적 인 얼룩. 최근의 작업은 SC에이 t에 한정되지 않을 수 있음을 강조지역 우와, 그리고이 사실에 단단히 혈관 (11)와 관련된 세포의 대부분 대기 인구로 뇌 전반에 걸쳐 현지화. SC에가 부상에 응답 동원 관측은 신경 퇴행성 질환 및 뇌졸중 (12, 13)에서 복구를 돕기 위해 회생 용으로 이들 세포를 이용할 수 있다는 가능성을 시사한다. 따라서 골아 세포, 연골 세포 및 지방 세포 (14)가 될 가능성이있는 혈관 주위 세포로 발견되는 중간 엽 줄기 세포 (MSC들)이 결합 조직의 치유에 재생하는 역할 달리 아니다. 그러나 NSCs의 일상적인 흡인 및 밀도 구배 원심 분리 기술에 의해 골수에서 중간 엽 줄기 세포와 동일하게하여 수확 할 수없고 이후자가 세포 기반 치료에 활용. 결과적으로, 그러한의 emb 유래 태아 NSCs의 또는 신경 전구체의 사용과 같은 셀의 다른 소스ryonic 줄기 세포가 광범위하게 성공 (15)의 학위를 변화와 동물의 질병과 부상의 모델을 탐구하고 있습니다. 체세포 소스를 이용하여 유도 만능 줄기 세포 기술은 제한된 가용성 및 배아 세포와 태아 조직 (16)의 사용에 대한 윤리적 문제를 극복하고, 응용 프로그램의 광범위한 치료에 유용 세포 기반 치료법을 생산하는 또 다른 잠재적 인 수단을 제공합니다. 그러나, 이러한 연구 결과의 임상 번역 (17, 18)뿐만 아니라, 규제 통관에 구불 구불 한 경로를 접근 SC 기반 치료와 함께 다양한 신경 학적 조건을 치료하는 투쟁에서와 같이, 어려운 작업이 될 입증되었습니다. 다른 방법으로, 특정 약물 치료의 도입은 NSC의 숫자를 조절하고 파킨슨 질환과 뇌졸중의 19 모델의 회복을 촉진 할 수있다. 전략은 효과적으로 이러한 세포를 조작하는 방법을 이해 할 수있는 어떤접근 체외 시스템이 필요합니다. NSCs의의 문화는 neurosphere를로 알려진 집계 문화, 또는 단일 층 8,20로도 수행 할 수 있습니다. 두 기술은 널리 다 능성 전구 물질의 확장을 제공 정의 배양 조건, 유사 분열 물질 소스로 표피 성장 인자 (EGF) 또는 기본 섬유 아세포 성장 인자 (bFGF의)의 예 사용의 설립을 허용, 사용되어왔다. neurosphere 배양 고립 셀 분류 클론 번식 능력을 연구에 더 적합 할 수 있지만, 시스템 (21)은 팽창시 세포의 혼합 집단을 생성하는 것으로 나타났다. 또, neurosphere를의 밀폐 구조는 세포의 약리 조작 비실용적하게하고, 이러한 요인이있을 수 영향 해석 인해 neurosphere 자체 내에 설립 미시로 혼동 할 수있다. 단층 배양, 다른 한편으로는, 일 수있다SC의 성장과 분화를 조절하고 특히이 세포 집단을 대상으로 새로운 화합물을 발견 할 수있는 기회를 여는 신호 전달 메커니즘을 탐구 할 수있는 강력한 도구를 제공하고, 작은 분자 라이브러리의 높은 처리량 화면에 사용. 결과적으로, 재현성 뇌 그 상이한 영역에서 성인 NSCs의 배양을 생성하는 기능은 중추 신경계 (CNS)의 개발 연구로부터 신규 한 재생 의료 방법을 탐구에 이르는 연구 광범위한 애플리케이션에 사용될 수있다 . 여기에 제시된 프로토콜은 해부 성인 쥐의 뇌에서 분리 CNS의 SC와의 차별화의 잠재력을 평가하는 방법을 보여줍니다.

<table border="0" cellpadding="0" cellspacing="0" width="596">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">6-well tissue culture dishes</td>
			<td style="width:95px;">BD Biosciences</td>
			<td style="width:119px;">353934</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="105">
			<td height="190" rowspan="1" style="height:190px;width:216px;">Poly-L-ornithine</td>
			<td rowspan="1" style="width:95px;">Sigma-Aldrich</td>
			<td rowspan="1" style="width:119px;">P-3655</td>
			<td style="width:167px;">
			5 mg/ml stock solution prepared in double distilled water (Stable for several months at -20 &#176;C).
			Working concentration 0.5 mg/ml in water (Stable for 1 month at 4 &#176;C).
			</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Fibronectin</td>
			<td style="width:95px;">R&#38;D Systems</td>
			<td style="width:119px;">1030-Fn</td>
			<td style="width:167px;">Do not agitate stock solution</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Filtration Apparatus</td>
			<td style="width:95px;">Corning Life Sciences</td>
			<td style="width:119px;">430769</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:216px;">DMEM/F-12</td>
			<td style="width:95px;">Mediatech</td>
			<td style="width:119px;">10-090-CV</td>
			<td style="width:167px;">See note below for complete N2 media preparation</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Apo-transferrin</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">T-2036</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Insulin</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">I-0516</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="68">
			<td height="68" style="height:68px;width:216px;">Putrescine</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">P-5780</td>
			<td style="width:167px;">1 M stock solution in ddH2O (Stable at 20 &#176;C for 6 months)</td>
		</tr>
		<tr height="68">
			<td height="68" style="height:68px;width:216px;">Sodium Selenite</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">S-5261</td>
			<td style="width:167px;">500 &#181;M stock solution in ddH2O (Stable at -20 &#176;C for 6 months)</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:216px;">Progesterone</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">P-8783</td>
			<td style="width:167px;">100 &#181;M stock solution in ethanol (Stable at -20&#176;C for 6 months)</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Penicillin/Streptomycin</td>
			<td style="width:95px;">Invitrogen</td>
			<td style="width:119px;">15140-122</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Phosphate Buffered Saline</td>
			<td style="width:95px;">Mediatech</td>
			<td style="width:119px;">21-040-CV</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Basic Fibroblast Growth Factor (bFGF)</td>
			<td style="width:95px;">R&#38;D Systems</td>
			<td style="width:119px;">233-FB</td>
			<td style="width:167px;">Working concentration 20 ng/ml&#160;</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Delta4 (Dll4)</td>
			<td style="width:95px;">R&#38;D Systems</td>
			<td style="width:119px;">1389-D4</td>
			<td style="width:167px;">Working concentration&#160; 500 ng/ml</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Angiopoetin 2 (Ang2)</td>
			<td style="width:95px;">R&#38;D Systems</td>
			<td style="width:119px;">623-AN</td>
			<td style="width:167px;">Working concentration&#160; 500 ng/ml</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">JAK Inhibitor</td>
			<td style="width:95px;">Calbiochem</td>
			<td style="width:119px;">420099</td>
			<td style="width:167px;">Working concentration 200 nM&#160;</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Bovine Serum Albumin</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">A-2058</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">15- and 50-ml Conical tubes</td>
			<td style="width:95px;">Corning Life Sciences</td>
			<td style="width:119px;">430053, 430829</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="243">
			<td colspan="4" height="243" style="height:243px;width:596px;">Other necessary items include: General dissection instruments, including razor blade and forceps, Adult rat (3-6 months old; Sprague-Dawley or Long Evans), CO2 intoxication chamber, Laminar flow hood for cell culture and incubator Incubator (humidified, 37 &#176;C, 5% CO2, 5% O2). Note: For complete N2 media preparation, to one bottle of DMEM/F-12 (500 ml) add 0.05 g of apotransferin, 0.0125 g of insulin (freshly predissolved in 1 ml of 10 mM NaOH), 50 &#956;l of putrescine, 30 &#956;l of sodium selenite, 100 &#956;l of progesterone stocks, and 5 ml of penicillin/streptomycin solution.&#160; Adjust pH to 7.2, if needed. Filter-sterilize and store at 4 &#176;C for up to 3 weeks and protect from light.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;"></td>
			<td style="width:95px;"></td>
			<td style="width:119px;"></td>
			<td>[header]</td>
		</tr>
		<tr height="22">
			<td colspan="2" height="22" style="height:22px;"></td>
			<td></td>
			<td></td>
			<td>Immunofluorescence Reagents Table</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:216px;">Paraformaldehyde</td>
			<td style="width:95px;">Electron Microscopy Sciences</td>
			<td style="width:119px;">15719</td>
			<td></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Normal Donkey Serum (NDS)</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">D-9663</td>
			<td></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Triton X-100</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">T-8787</td>
			<td></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">4,6-Diamidino-2-phenylindole (DAPI)</td>
			<td style="width:95px;">Sigma-Aldrich</td>
			<td style="width:119px;">D-8417</td>
			<td style="width:167px;">5 mg/ml stock solution in methanol</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;"></td>
			<td></td>
			<td></td>
			<td>[header]</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;"></td>
			<td style="width:95px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Primary Antibody Table</td>
		</tr>
		<tr height="21">
			<td height="43" rowspan="1" style="height:43px;width:216px;">Nestin</td>
			<td rowspan="1" style="width:95px;">Chemicon</td>
			<td rowspan="1" style="width:119px;">MAB353</td>
			<td style="width:167px;">
			Dilution Factor: 1:400
			Species: Mouse IgG1
			</td>
		</tr>
		<tr height="21">
			<td height="43" rowspan="1" style="height:43px;width:216px;">Hes3</td>
			<td rowspan="1" style="width:95px;">Santa Cruz</td>
			<td rowspan="1" style="width:119px;">sc-25393</td>
			<td style="width:167px;">
			Dilution Factor: 1:100
			Species: Rabbit IgG
			</td>
		</tr>
		<tr height="21">
			<td height="43" rowspan="1" style="height:43px;width:216px;">Sox2</td>
			<td rowspan="1" style="width:95px;">R&#38;D Systems</td>
			<td rowspan="1" style="width:119px;">MAB2018&#160;</td>
			<td style="width:167px;">
			Dilution Factor: 1:100
			Species: Mouse IgG2a
			</td>
		</tr>
		<tr height="21">
			<td height="43" rowspan="1" style="height:43px;width:216px;">CNPase</td>
			<td rowspan="1" style="width:95px;">Chemicon</td>
			<td rowspan="1" style="width:119px;">MAB326</td>
			<td style="width:167px;">
			Dilution Factor: 1:200
			Species: Mouse IgG1
			</td>
		</tr>
		<tr height="21">
			<td height="43" rowspan="1" style="height:43px;width:216px;">&#946;-tubulin III (TUJ1)</td>
			<td rowspan="1" style="width:95px;">R&#38;D Systems</td>
			<td rowspan="1" style="width:119px;">MAB1195</td>
			<td style="width:167px;">
			Dilution Factor: 1:500
			Species: Mouse IgG2a
			</td>
		</tr>
		<tr height="21">
			<td height="43" rowspan="1" style="height:43px;width:216px;">Glial Fibrillary Acidic Protein (GFAP)</td>
			<td rowspan="1" style="width:95px;">Dako North America</td>
			<td rowspan="1" style="width:119px;">Z0334</td>
			<td style="width:167px;">
			Dilution Factor 1:500
			Species: Rabbit
			</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;"></td>
			<td></td>
			<td></td>
			<td>[header]</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;"></td>
			<td style="width:95px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Secondary Antibody Table</td>
		</tr>
		<tr height="21">
			<td height="64" rowspan="1" style="height:64px;width:216px;">Alexa&#160;&#160; 568</td>
			<td rowspan="1" style="width:95px;">Invitrogen</td>
			<td rowspan="1" style="width:119px;">A-21124</td>
			<td style="width:167px;">
			Dilution Factor: 1:200
			Species: Goat anti Mouse IgG1
			</td>
		</tr>
		<tr height="21">
			<td height="64" rowspan="1" style="height:64px;width:216px;">Alexa 488</td>
			<td rowspan="1" style="width:95px;">Invitrogen</td>
			<td rowspan="1" style="width:119px;">A-21131</td>
			<td style="width:167px;">
			Dilution Factor: 1:200
			Species: Goat anti Mouse IgG2a
			</td>
		</tr>
		<tr height="21">
			<td height="64" rowspan="1" style="height:64px;width:216px;">Cy5</td>
			<td rowspan="1" style="width:95px;">Jackson ImmunoResearch</td>
			<td rowspan="1" style="width:119px;">59883</td>
			<td style="width:167px;">
			Dilution Factor: 1:200
			Species: Goat anti Rabbit
			</td>
		</tr>
	</tbody>
</table>

growing neural stem cells from conventional and nonconventional regions of the adult rodent brain

최근 연구는 중추 신경계 (CNS) 재생 및 종양 줄기 세포의 인구 (SC에) 성인 뇌 내 거주자를 포함 것을 보여줍니다. 그러나 메커니즘이 정상적으로 정지 세포가 적절한 신경망의 기능뿐만 아니라, 신경 퇴행성 프로세스의 부상 및 완화 회복에 자신의 역할을 보장하기 위해 고용은 조금 이해된다. 이러한 세포는 혈관 및 면역 조절 성 시스템 모두로부터 신호들을 포함한 서스테인 환경을 제공 「틈새」이라고도 영역에 상주. 알 수없는 요인을 제외 정의 배양 조건에서 CNS의 SC와의 분리, 유지 보수 및 차별화, 따라서 자신의 생체 내 행동에 대한 통찰력을 제공하고, 약물 또는 유전자에 의한 치료에 액세스 할 수 있습니다. 여기에서 우리는 성인 뇌의 특정 영역에서 CNS의 SC와의 문화를 생성하는 방법에 대한 자세한 정보를 제공하고 자신의 개발을 평가하는 접근신경 세포, 성상 세포, 그리고 체외에서 희소 돌기 아교 세포에 ifferentiation 가능성. 이 기술은 개별 SC에 자신의 자손을 연구하기 위해 시각화 할 수있는 단일 층 문화로 균일 한 세포 인구를 산출한다. 게다가, 다른 동물 모델 시스템과 임상 샘플을 가로 질러인가 될 수 손상된 성인 신경계 재생 반응을 예측하기 이전에 사용된다.

신경 줄기 세포를 수확되는 관심 영역을 확인하는 것은 중요한 첫 단계이며 그것은 세포의 합류 판을 얻​​는 데 걸리는 시간의 양을 정의 할 것이다. 예를 들어, SVZ는 고전 신경성 영역이며, 따라서 신경 줄기 세포의 상대적으로 높은 비율을 갖는다. 그러나, 여기에 제시된 기술은 종종 성인기 강력한 신경성 잠재력을 갖는 것으로 인식되지 않는 다른 영역으로 사용될 수있다. 이 프로토콜의 목?...

Watch this Scientific Journal Video about Growing Neural Stem Cells from Conventional and Nonconventional Regions of the Adult Rodent Brain at JoVE.com

Growing Neural Stem Cells from Conventional and Nonconventional Regions of the Adult Rodent Brain

성인의 뇌에서 수확 한 신경 줄기 세포는 신경 시스템 개발의 기초 연구에서 재생 의학의 잠재적 인 임상 응용 프로그램을 탐험에 이르기까지 다양한 애플리케이션에 활용 증가하고있다. 이 실험 결과를 소리를하는 것이 중요합니다 이러한 세포를 성장하는 데 사용되는 분리 및 배양 조건에서 엄격한 통제를한다.

성인 쥐의 뇌의 기존 및 비 인습적 인 지역에서 신경 줄기 세포 성장

Recent work demonstrates that central nervous system (CNS) regeneration and tumorigenesis involves populations of stem cells (SCs) resident within the ...

growing-neural-stem-cells-from-conventional-nonconventional-regions

Research

JoVE Journal

Neuroscience

12.1K 조회수.  University of Dresden.  성인의 뇌에서 수확 한 신경 줄기 세포는 신경 시스템 개발의 기초 연구에서 재생 의학의 잠재적 인 임상 응용 프로그램을 탐험에 이르기까지 다양한 애플리케이션에 활용 증가하고있다. 이 실험 결과를 소리를하는 것이 중요합니다 이러한 세포를 성장하는 데 사용되는 분리 및 배양 조건에서 엄격한 통제를한다. 

비디오: Growing Neural Stem Cells from Conventional and Nonconventional Regions of the Adult Rodent Brain

Growth and Differentiation of Adult Hippocampal Arctic Ground Squirrel Neural Stem Cells

Arctic ground squirrels (Urocitellus parryii, AGS) are unique in their ability to hibernate with a core body temperature near or below freezing 1. These animals also resist ischemic injury to the brain in vivo 2,3 and oxygen-glucose deprivation in vitro 4,5. These unique qualities provided the impetus to isolate AGS neurons to examine inherent neuronal characteristics that could account for the capacity of AGS neurons to resist injury and cell death caused by ischemia and extremely cold temperatures. Identifying proteins or gene targets that allow for the distinctive properties of these cells could aid in the discovery of effective therapies for a number of ischemic indications and for the study of cold tolerance. Adult AGS hippocampus contains neural stem cells that continue to proliferate, allowing for easy expansion of these stem cells in culture. We describe here methods by which researchers can utilize these stem cells and differentiated neurons for any number of purposes. By closely following these steps the AGS neural stem cells can be expanded through two passages or more and then differentiated to a culture high in TUJ1-positive neurons (~50%) without utilizing toxic chemicals to minimize the number of dividing cells. Ischemia induces neurogenesis 6 and neurogenesis which proceeds via MEK/ERK and PI3K/Akt survival signaling pathways contributes to ischemia resistance in vivo7 and in vitro 8 (Kelleher-Anderson, Drew et al., in preparation). Further characterization of these unique neural cells can advance on many fronts, using some or all of these methods.

Neural stem cells were prepared from the hippocampus of adult non-hibernating yearling Arctic ground squirrels (AGS). These neural stem cells can be expanded through numerous passages, differentiated and maintained as a nearly 50:50 neuron to glial culture.

성인 Hippocampal 북극 그라운드 다람쥐 신경 줄기 세포의 성장과 분화

Arctic ground squirrels (Urocitellus parryii, AGS) are unique in their ability to hibernate with a core body temperature near or below freezing 1. These ...

연구

신경과학

Isolation and Expansion of the Adult Mouse Neural Stem Cells Using the Neurosphere Assay

Isolation and expansion of the putative neural stem cells (NSCs) from the adult murine brain was first described by Reynolds and Weiss in 1992 employing a chemically defined serum-free culture system known as the neurosphere assay (NSA). In this assay, the majority of differentiated cell types die within a few days of culture but a small population of growth factor responsive precursor cells undergo active proliferation in the presence of epidermal growth factor (EGF) and/ basic fibroblastic growth factor (bFGF). These cells form colonies of undifferentiated cells called neurospheres, which in turn can be subcultured to expand the pool of neural stem cells. Moreover, the cells can be induced to differentiate, generating the three major cell types of the CNS i.e. neurons, astrocytes, and oligodendrocytes. This assay provides an invaluable tool to supply a consistent, renewable source of undifferentiated CNS precursors, which could be used for in vitro studies and also for therapeutic purposes.
This video demonstrates the NSA method to generate and expand NSCs from the adult mouse periventricular region, and provides technical insights to ensure one can achieve reproducible neurosphere cultures. The procedure includes harvesting the brain from the adult mouse, micro-dissection of the periventricular region, tissue preparation and culture in the NSA. The harvested tissue is first chemically digested using trypsin-EDTA and then mechanically dissociated in NSC medium to achieve a single cell suspension and finally plated in the NSA. After 7-10 days in culture, the resulting primary neurospheres are ready for subculture to reach the amount of cells required for future experiments.

This video protocol demonstrates the neurosphere assay method to generate and expand neural stem cells from the adult mouse periventricular region, and provides technical insights to ensure one can achieve reproducible neurosphere cultures.

절연 및 Neurosphere 어세를 사용하여 성인 마우스 신경 줄기 세포의 확장

Isolation and expansion of the putative neural stem cells (NSCs) from the adult murine brain was first described by Reynolds and Weiss in 1992 employing a ...

Isolation and Culture of Neural Crest Cells from Embryonic Murine Neural Tube

The embryonic neural crest (NC) is a multipotent progenitor population that originates at the dorsal aspect of the neural tube, undergoes an epithelial to mesenchymal transition (EMT) and migrates throughout the embryo, giving rise to diverse cell types 1-3. NC also has the unique ability to influence the differentiation and maturation of target organs4-6. When explanted in vitro, NC progenitors undergo self-renewal, migrate and differentiate into a variety of tissue types including neurons, glia, smooth muscle cells, cartilage and bone. 
NC multipotency was first described from explants of the avian neural tube7-9. In vitro isolation of NC cells facilitates the study of NC dynamics including proliferation, migration, and multipotency. Further work in the avian and rat systems demonstrated that explanted NC cells retain their NC potential when transplanted back into the embryo10-13. Because these inherent cellular properties are preserved in explanted NC progenitors, the neural tube explant assay provides an attractive option for studying the NC in vitro. 
To attain a better understanding of the mammalian NC, many methods have been employed to isolate NC populations. NC-derived progenitors can be cultured from post-migratory locations in both the embryo and adult to study the dynamics of post-migratory NC progenitors11,14-20, however isolation of NC progenitors as they emigrate from the neural tube provides optimal preservation of NC cell potential and migratory properties13,21,22. Some protocols employ fluorescence activated cell sorting (FACS) to isolate a NC population enriched for particular progenitors11,13,14,17. However, when starting with early stage embryos, cell numbers adequate for analyses are difficult to obtain with FACS, complicating the isolation of early NC populations from individual embryos. Here, we describe an approach that does not rely on FACS and results in an approximately 96% pure NC population based on a Wnt1-Cre activated lineage reporter23. 
The method presented here is adapted from protocols optimized for the culture of rat NC11,13. The advantages of this protocol compared to previous methods are that 1) the cells are not grown on a feeder layer, 2) FACS is not required to obtain a relatively pure NC population, 3) premigratory NC cells are isolated and 4) results are easily quantified. Furthermore, this protocol can be used for isolation of NC from any mutant mouse model, facilitating the study of NC characteristics with different genetic manipulations. The limitation of this approach is that the NC is removed from the context of the embryo, which is known to influence the survival, migration and differentiation of the NC2,24-28.

Isolation of embryonic neural crest from the neural tube facilitates the use of in vitro methods for studying migration, self-renewal, and multipotency of neural crest.

배아 Murine 신경 튜브에서 신경 크레스트 세포의 분리 및 문화

The embryonic neural crest (NC) is a multipotent progenitor population that originates at the dorsal aspect of the neural tube, undergoes an epithelial to ...

One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice

The neurosphere assay and the adherent monolayer culture system are valuable tools to determine the potential (proliferation or differentiation) of adult neural stem cells in vitro. These assays can be used to compare the precursor potential of cells isolated from genetically different or differentially treated animals&#160;to determine the effects of exogenous factors on neural precursor cell proliferation and differentiation and to generate neural precursor cell lines that can be assayed over continuous passages. The neurosphere assay is traditionally used for the post-hoc identification of stem cells, primarily due to the lack of definitive markers with which they can be isolated from primary tissue&#160;and has the major advantage of giving a quick estimate of precursor cell numbers in brain tissue derived from individual animals. Adherent monolayer cultures, in contrast, are not traditionally used to compare proliferation between individual animals, as each culture is generally initiated from the combined tissue of between 5-8 animals. However, they have the major advantage that, unlike neurospheres, they consist of a mostly homogeneous population of precursor cells and are useful for following the differentiation process in single cells. Here, we describe, in detail, the generation of neurosphere cultures and, for the first time, adherent cultures from individual animals. This has many important implications including paired analysis of proliferation and/or differentiation potential in both the subventricular zone (SVZ) and dentate gyrus (DG) of treated or genetically different mouse lines, as well as a significant reduction in animal usage.

Here we describe a detailed protocol for the simultaneous generation of neural precursor cell cultures, as either adherent monolayers or neurospheres, from the subventricular zone and dentate gyrus of individual adult mice.

하나의 마우스, 두 문화 : 분리 및 성인 신경의 문화가 개인 마우스의 두 신경 인성 영역에서 줄기 세포

The neurosphere assay and the adherent monolayer culture system are valuable tools to determine the potential (proliferation or differentiation) of adult ...

Feeder-free Derivation of Neural Crest Progenitor Cells from Human Pluripotent Stem Cells

Human pluripotent stem cells (hPSCs) have great potential for studying human embryonic development, for modeling human diseases in the dish and as a source of transplantable cells for regenerative applications after disease or accidents. Neural crest (NC) cells are the precursors for a large variety of adult somatic cells, such as cells from the peripheral nervous system and glia, melanocytes and mesenchymal cells. They are a valuable source of cells to study aspects of human embryonic development, including cell fate specification and migration. Further differentiation of NC progenitor cells into terminally differentiated cell types offers the possibility to model human diseases in vitro, investigate disease mechanisms and generate cells for regenerative medicine. This article presents the adaptation of a currently available in vitro differentiation protocol for the derivation of NC cells from hPSCs. This new protocol requires 18 days of differentiation, is feeder-free, easily scalable and highly reproducible among human embryonic stem cell (hESC) lines as well as human induced pluripotent stem cell (hiPSC) lines. Both old and new protocols yield NC cells of equal identity.

Neural crest (NC) cells derived from human pluripotent stem cells (hPSC) have great potential for modeling human development and disease and for cell replacement therapies. Here, a feeder-free adaptation of the currently widely used in vitro differentiation protocol for the derivation of NC cells from hPSCs is presented.

인간 다 능성에서 신경 능선 세포의 전구 세포의 공급이없는 유도는 줄기 세포

Human pluripotent stem cells (hPSCs) have great potential for studying human embryonic development, for modeling human diseases in the dish and as a ...

Cell Sorting of Neural Stem and Progenitor Cells from the Adult Mouse Subventricular Zone and Live-imaging of their Cell Cycle Dynamics

Neural stem cells (NSCs) in the subventricular zone of the lateral ventricles (SVZ) sustain olfactory neurogenesis throughout life in the mammalian brain. They successively generate transit amplifying cells (TACs) and neuroblasts that differentiate into neurons once they integrate the olfactory bulbs. Emerging fluorescent activated cell sorting (FACS) techniques have allowed the isolation of NSCs as well as their progeny and have started to shed light on gene regulatory networks in adult neurogenic niches. We report here a cell sorting technique that allows to follow and distinguish the cell cycle dynamics of the above-mentioned cell populations from the adult SVZ with a LeX/EGFR/CD24 triple staining. Isolated cells are then plated as adherent cells to explore in details their cell cycle progression by time-lapse video microscopy. To this end, we use transgenic Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) mice in which cells are red-fluorescent during G1 phase due to a G1 specific red-Cdt1 reporter. This method has recently revealed that proliferating NSCs progressively lengthen their G1 phase during aging, leading to neurogenesis impairment. This method is easily transposable to other systems and could be of great interest for the study of the cell cycle dynamics of brain cells in the context of brain pathologies.

We report a Fluorescent Activated Cell Sorting (FACS)-based method to isolate neural stem cells (NSCs) and their progeny from the subventricular zone (SVZ) of the adult mouse brain. Applied to Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) transgenic mice, it allows the study of cell cycle progression by live imaging.

셀 성인 마우스 뇌실 영역에서 신경 줄기 및 전구 세포의 정렬과 세포주기 역학의 라이브 이미징

Neural stem cells (NSCs) in the subventricular zone of the lateral ventricles (SVZ) sustain olfactory neurogenesis throughout life in the mammalian brain. ...

Isolation and Culture of Adult Neural Stem Cells from the Mouse Subcallosal Zone

Adult neural stem cells (aNSCs) can be used for the regeneration of damaged brain tissue. NSCs have the potential for differentiation and proliferation into three types of cells: neurons, astrocytes, and oligodendrocytes. Identifying aNSC-derived regions and characterizing the aNSC properties are critical for the potential use of aNSCs and for the elucidation of their role in neural regeneration. The subcallosal zone (SCZ), located between white matter and the hippocampus, has recently been reported to contain aNSCs and continuously give rise to neuroblasts. A low percentage of aNSCs from the SCZ is differentiated into neurons; most cells are differentiated into glial cells, such as oligodendrocytes and astrocytes. These cells are suggested to have a therapeutic potential for traumatic cortical injury. This protocol describes in detail the process to generate SCZ-aNSCs from an adult mouse brain. A brain matrix with intervals of 1 mm is used to obtain the SCZ-containing coronal slices and to precisely dissect the SCZ from the whole brain. The SCZ sections are initially subjected to a neurosphere culture. A well-developed culture system allows for the verification of their characteristics and can increase research on NSCs. A neurosphere culture system provides a useful tool for determining proliferation and collecting the genuine NSCs. A monolayer culture is also an in vitro system to assay proliferation and differentiation. Significantly, this culture system provides a more homogenous environment for NSCs than the neurosphere culture system. Thus, using a discrete brain region, these culture systems will be helpful for expanding our knowledge about aNSCs and their applications for therapeutic uses.

Establishing culture systems for the expansion of adult neural stem cells (aNSCs) allows for the examination and application of aNSCs for therapy. The subcallosal zone (SCZ) has recently been recognized as a novel neuroblast-forming region in adult mice. Here, methods for the isolation, expansion, and differentiation of SCZ-aNSCs are described.

마우스 뇌량 밑 영역에서의 분리 및 성인 신경 줄기 세포의 문화

Adult neural stem cells (aNSCs) can be used for the regeneration of damaged brain tissue. NSCs have the potential for differentiation and proliferation ...

Use of Synaptic Zinc Histochemistry to Reveal Different Regions and Laminae in the Developing and Adult Brain

Characterization of anatomical and functional brain organization and development requires accurate identification of distinct neural circuits and regions in the immature and adult brain. Here we describe a zinc histochemical staining procedure that reveals differences in staining patterns among different layers and brain regions. Others have utilized this procedure not only to reveal the distribution of zinc-containing neurons and circuits in the brain, but also to successfully delineate areal and laminar boundaries in the developing and adult brain in several species. Here we illustrate this staining procedure with images from developing and adult ferret brains. We reveal a zinc-staining pattern that serves as an anatomical marker of areas and layers, and can be reliably used to distinguish visual cortical areas in the developing and adult visual cortex. The main goal of this protocol is to present a histochemical method that allows the accurate identification of layers and regions in the developing and adult brain where other methods fail to do so. Secondarily, in conjunction with densitometric image analysis, this method allows one to assess the distribution of synaptic zinc to reveal potential changes throughout development. This protocol describes in detail the reagents, tools, and steps necessary to successively stain frozen brain sections. Although this protocol is described using ferret brain tissue, it can easily be adapted for use in rodents, cats, or monkeys as well as in other brain regions.

We describe a histochemical procedure that reveals characteristic laminar and areal zinc staining patterns in different brain regions. The zinc-staining pattern may be used in conjunction with other anatomical markers to reliably distinguish layers and regions in the developing and adult brain.

시 냅 스 아연 Histochemistry 다른 지역과 개발 및 성인 두뇌에서 하기의 사용

Characterization of anatomical and functional brain organization and development requires accurate identification of distinct neural circuits and regions ...

Collection of Frozen Rodent Brain Regions for Downstream Analyses

As our understanding of neurobiology has progressed, molecular analyses are often performed on small brain areas such as the medial prefrontal cortex (mPFC) or nucleus accumbens. The challenge in this work is to dissect the correct area while preserving the microenvironment to be examined. In this paper, we describe a simple, low-cost method using resources readily available in most labs. This method preserves nucleic acid and proteins by keeping the tissue frozen throughout the process. Brains are cut into 0.5&#8211;1.0 mm sections using a brain matrix and arranged on a frozen glass plate. Landmarks within each section are compared to a reference, such as the Allen Mouse Brain Atlas, and regions are dissected using a cold scalpel or biopsy punch. Tissue is then stored at -80 &#176;C until use. Through this process rat and mouse mPFC, nucleus accumbens, dorsal and ventral hippocampus and other regions have been successfully analyzed using qRT-PCR and Western assays. This method is limited to brain regions that can be identified by clear landmarks.

This procedure describes the collection of discrete frozen brain regions to obtain high-quality protein and RNA using inexpensive and commonly available tools.

다운스트림 분석을 위한 냉동 설치류 뇌 영역 모음

As our understanding of neurobiology has progressed, molecular analyses are often performed on small brain areas such as the medial prefrontal cortex ...

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to intracranial delivery, intra-arterial administration of NSCs is less invasive and produces a more diffuse distribution of NSCs within the brain parenchyma. Further, intra-arterial delivery allows the first-pass effect in the brain circulation, lessening the potential for trapping of cells in peripheral organs, such as liver and spleen, a complication associated with peripheral injections. Here, we detail the methodology, in both mice and rats, for delivery of NSCs through the common carotid artery (mouse) or external carotid artery (rat) to the ipsilateral hemisphere after an ischemic stroke. Using GFP-labeled NSCs, we illustrate the widespread distribution achieved throughout the rodent ipsilateral hemisphere at 1 d, 1 week and 4 weeks after postischemic delivery, with a higher density in or near the ischemic injury site. In addition to long-term survival, we show evidence of differentiation of GFP-labeled cells at 4 weeks. The intra-arterial delivery approach described here for NSCs can also be used for administration of therapeutic compounds, and thus has broad applicability to varied CNS injury and disease models across multiple species.

A method for delivering neural stem cells, adaptable for injecting solutions or suspensions, through the common carotid artery (mouse) or external carotid artery (rat) after ischemic stroke is reported. Injected cells are distributed broadly throughout the brain parenchyma and can be detected up to 30 d after delivery.

쥐와 마우스 두뇌에 신경 줄기 세포의 동맥 내 납품: 대뇌 허혈에 응용 프로그램

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...