Name: isolation of infiltrating leukocytes from mouse skin using enzymatic digest and gradient separation
Uploaded: 2016-01-25T18:00:00
Description: Watch this Scientific Journal Video about Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation at JoVE.com

국립 보건원 (DC에 NIH R15 NS067536-01A1), 국립 Vulvodynia 협회 (DC에 수상), 그리고 매 켈리 스터 대학교는이 작업을 지원했다. 인증 기관은 아놀드와 로벨 베크 재단 자금 매 켈리 스터 대학교 베크 학자 프로그램에서 교제를 받았다. BTF 및 TM은 JDRF 2-2011-662 지원합니다. 우리는 기술적 조언 박사 제이슨 Schenkel 박사 줄리아 루이스 감사, 그들의 도움과 지원에 대한 Chatterjea 랩의 모든 현재의 전 멤버.

주 : 통상적으로 음식과 물을 무료로 이용할 수와 보관 8~12주 세 여자 ND4 스위스 웹스터 마우스는, 이러한 연구에 사용 하였다 (B13S1)에 사용되는 실험 프로토콜은 매 켈리 스터 대학교의 IACUC에 의해 승인되었다.. 옥사 졸론 1. 과민성 및 도전 <ol><li> 날 0 <ol><li> 4 L 뚜껑 유리 항아리의 하단에 메쉬 아래에 배치 흡수성 수건에 3 ㎖ 이소 플루 란을 추가하여 마취 챔버를 준비합니다. 피사...

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이러한 아토피 성 피부염이나 건선과 같은 피부 질환의 설치류 모델에서 피부에 상주하는 백혈구의 변화를 특성화하는 염증 세포의 유입과 질병 병리 사이의 기계적 연결을 이해하는데 중요하다. 여기에서 우리는 대부분의 생물 의학 연구 실험실에서 발견되는 기본 장비로 피부 조직에서 백혈구를 분리하는 경제적 인 방법을 설명합니다. 이 상대적으로 빠른 기술은 실험실 벤치에서 시간에 손?...

접촉 성 피부염, 습진, 건선, 봉와직염, 비 흑색 종 피부암에 곰팡이 감염 및 농양에 이르기까지 피부 상태는 전세계 50 가장 널리 퍼진 질병들로 발견되었고 2010 1 치명적 질병의 네 번째 선도적 인 글로벌 원인. 따라서, 다양한 피부 병변을 기본 분자 및 세포 메커니즘의 조사 연구에 필요한 활성 영역입니다. 설치류 모델은 아토피 성 피부염 (2), (3), 건선, 또는 포도상 구균 감염 4 같은 염증성 피부 질환의 이해에 도움이 현저하게되어왔다. 마우스 피부 조직의 효소 적 소화, 저렴하고 효율적이며 간단한 프로토콜 나은 피부 질환의 병태 생리를 이해 하류 다양한 용도에 사용할 수있는 세포의 제제를 제공 할 수있다. 여기서, 간단하고 경제적 인 방법은 마우스 피부 효소에 대해 기재되어 다이제스트조직 및 세포 배양, 생체 대체 전송에 사용할 수있는 피부 침투 백혈구의 분리, 유세포 분석 및 분류 또는 유전자 발현 연구를 흐른다. 이 절차의 목적은 통상적으로 전체적인 맞춤 시약 키트 및 기계적 dissociators와 연관된 비용을 최소화하면서 높은 세포 생존율과 피부 백혈구 침윤의 단일 세포 현탁액을 제조하는 공정이다. 기존의 피부 조직 해리 방법 5-7 낮은 세포 생존 및 표면 마커의 무결성 결과, 또는 사용자 정의 효소 키트와 고가의 조직 해리 기계 8-11 필요할 수 있습니다. 마우스 귀 피부 조직의 소화가 합리적으로, 12 ~ 13 유행 높은 각화 피부 조직을 소화하는 동안 (예를 들면 측면에서) 비 세포 파편 많은 양의 오염 된 세포 준비가 발생할 수 있습니다. 최근의 연구에서, 자이드와 동료가 2.5 mg을 90 분 동안 마우스 측면 피부를 소화 / ㎖ 디스 파제, FOLLO3 ㎎ / ㎖의 콜라게나 제 (7)에 의해 45 분 결혼. 또 다른 연구에서, 이들 연구자들은 트립신 / EDTA, 콜라겐 III의 사용을 포함하는, 2.5 시간의 조합으로 여러 소화 배양을 사용하고, (5) 디스 파제. 다른 제조업체의 트립신으로 처리 잴 포유 동물 세포에서 14-15 세포 표면 단백질의 무결성에 영향을 미치는 것으로 밝혀졌다 트립신의 사용은 피부 효소 소화에 권장되지 않는다. 또한, 디스 파제는 CD4 및 CD8α T 세포의 증식 능력에 심각한 영향을 미칠 수 및 CD62L (16)와 같은 일반적인 T 세포 활성화 마커를 포함, 최소 20 분자의 표면 풍요 로움에 영향을 미칩니다. 다른 프로토콜은 소화 매체 (6)에 RPMI 1640을 사용합니다. 그러나, RPMI에 Mg2 +, Ca2 + 등의 존재는 광범위한 세포 응집 (17)가 발생할 수있다. 조직 분리를위한 이상적인 프로토콜은 높은 세포 생존 능력, 낮은 목표로한다세포 응집, 최소한의 손상의 수준은 표면 단백질을 세포합니다. 고품질 림프절 간질 세포 제제는 짧은 효소 배양, 칼슘 및 마그네슘이없는 배지 2+ 프로토콜을 사용하여 수행하고, 트립신을 피하고 18 디스 파제되었다. 그러나, 이러한 유형의 프로토콜은 전체 마우스 피부의 해리에 대해 확립되지 않았다. 여기에서, 프로토콜은, 해리 분리 및 알레르겐 - 도전 마우스 옆구리 피부에서 피부 백혈구 침윤을 풍부하게 기재되어있다. 간략하게, 절제된 피부 소화 조직을 연화 초과 각질이나 지방 조직을 제거하기 위해 1 시간 동안 10 % 소 태아 혈청 행크의 균형 잡힌 염 용액 (HBSS)로 미리 배양된다. 이것은 0.7 mg을 함께 30 분간 효소 분해 단계 뒤에 / ㎖의 콜라게나 D. 콜라게나 D 그것을 만드는 최소 세포 표면 마커의 농도에 대한 영향, 및 시험관 (16, 18)에서 T 세포 증식에는 영향을주지표면 단백질의 특성을 포함하는 애플리케이션에 매우 적합하다. 효소 분해 후, 불연속 밀도 구배 원심 분리는 단일 세포 현탁액으로부터 상피 세포 및 잔해를 제거하고, 조혈 세포 농축 하였다. 중요한 것은,이 절차는 비싼 열 기반 자기 세포 분리 시약 및 티슈 기계 8-11 해리를 방지하고 기본 생의학 연구소에 위치한 재료 및 장비로 수행 될 수있다. 여기에서이 프로토콜 (19 각색) 이전에 감응 ND4 스위스 쥐 합텐 옥사 졸론 (황소)와 측면 피부에서 도전 세번 백혈구를 분리하는데 사용 하였다. 세포는 파라 메트릭 멀티 - 유동 세포 계측법을 사용하여 분석 하였다. 이 기술은 최소한의 파편과 계측법 영향 SK에 T 림프구와 호중구의 침윤을 측정하기 위해 파라 메트릭 다중 흐름으로 분석 하였다 격리 림프구의&gt; 95 %의 생존율을 가진 세포 현탁액을 수득에서.

<table border="0" cellpadding="0" cellspacing="0" width="1213">
	<tbody>
		<tr height="26">
			<td height="26" style="height:26px;width:457px;">HBSS with phenol red, without calcium, without magnesium, liquid</td>
			<td style="width:179px;">Sigma Aldrich</td>
			<td style="width:181px;">55021C</td>
			<td style="width:396px;">Keep sterile until day of usage.</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">HEPES, &#8805;99.5% (titration)</td>
			<td style="width:179px;">Sigma Aldrich</td>
			<td style="width:181px;">H3375</td>
			<td style="width:396px;"></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:457px;">EDTA disodium salt solution</td>
			<td style="width:179px;">Sigma Aldrich</td>
			<td style="width:181px;">E7889</td>
			<td style="width:396px;">Keep sterile until day of usage.</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:457px;">Fetal Bovine Serum, USDA, Heat Inactivated, Premium Select</td>
			<td style="width:179px;">MidSci</td>
			<td style="width:181px;">S01520HI</td>
			<td style="width:396px;">Keep sterile until day of usage.</td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:457px;">Percoll, pH 8.5-9.5 (25&#176;C)</td>
			<td style="width:179px;">Sigma Aldrich</td>
			<td style="width:181px;">P1644</td>
			<td style="width:396px;">Keep sterile. Percoll needs to be made isotonic with sterile 10X PBS prior to use.</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:457px;">Trimmer Combo Kit</td>
			<td style="width:179px;">Kent Scientific</td>
			<td style="width:181px;">CL9990-1201&#160;</td>
			<td style="width:396px;">Use the larger trimmer for shaving the flank and back.</td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:457px;">4-Ethoxymethylene-2-phenyl-2-oxazolin-5-one</td>
			<td style="width:179px;">Sigma Aldrich</td>
			<td style="width:181px;">E0753-10G</td>
			<td style="width:396px;">Dissolve in 100% EtOH at 50&#176;C for 15 minutes on a rotating plate.</td>
		</tr>
		<tr height="46">
			<td height="46" style="height:46px;width:457px;">Purified anti-mouse CD16/CD32 (2.4G2)</td>
			<td style="width:179px;">Tonbo Biosciences</td>
			<td style="width:181px;">70-0161</td>
			<td style="width:396px;">Antibody used to block non-specific antibody binding during antibody staining for flow cytometry</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:457px;">anti-CD3e-BV650 (145-2C11)</td>
			<td style="width:179px;">Becton Dickinson</td>
			<td style="width:181px;">564378</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-CD4-APC-eFluor780 (RM4-5)</td>
			<td style="width:179px;">eBioscience</td>
			<td style="width:181px;">47-0042-80</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-CD8a-BV785 (53-6.7)</td>
			<td style="width:179px;">BioLegend</td>
			<td style="width:181px;">100749</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-CD11b-eFluor450 (M1/70)</td>
			<td style="width:179px;">eBioscience</td>
			<td style="width:181px;">48-0112-80</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-CD11c-eFluor450 (N418)</td>
			<td style="width:179px;">eBioscience</td>
			<td style="width:181px;">48-0114-80</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-CD44-BV711 (IM7)</td>
			<td style="width:179px;">Becton Dickinson</td>
			<td style="width:181px;">563971</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-CD45-FITC (30-F11)</td>
			<td style="width:179px;">Tonbo Biosciences</td>
			<td style="width:181px;">35-0451-U025</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-CD45R-eFluor450 (RA3-6B2)</td>
			<td style="width:179px;">eBioscience</td>
			<td style="width:181px;">48-0452-80</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-CD62L-PerCP-Cy5.5 (MEL-14)</td>
			<td style="width:179px;">BioLegend</td>
			<td style="width:181px;">104431</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">anti-Ly-6G/Ly-6C (Gr-1)-PE (RB6-8C5)</td>
			<td style="width:179px;">BioLegend</td>
			<td style="width:181px;">108407</td>
			<td style="width:396px;">Antibody used to stain cells for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">Ghost Dye-BV510</td>
			<td style="width:179px;">Tonbo Biosciences</td>
			<td style="width:181px;">13-0870-T100</td>
			<td style="width:396px;">Viability dye for flow cytometry</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:457px;">LSR Fortessa</td>
			<td style="width:179px;">Becton Dickinson</td>
			<td style="width:181px;">N/A</td>
			<td style="width:396px;">Cell analyzer (18 parameters)</td>
		</tr>
		<tr height="96">
			<td height="96" style="height:96px;width:457px;">Collagenase D from Clostridium histolyticum</td>
			<td style="width:179px;">Roche Applied Science</td>
			<td style="width:181px;">11088858001</td>
			<td style="width:396px;">Aliquot lyophilized enzyme at 5 mg/mL in HBSS with phenol red, without calcium, and without magnesium into 1 mL aliquots.&#160; Store immediately at -20&#176;C for up to six months.</td>
		</tr>
		<tr height="92">
			<td height="92" style="height:92px;width:457px;">ND4 Swiss female mice</td>
			<td style="width:179px;">Harlan</td>
			<td style="width:181px;">0-32</td>
			<td style="width:396px;">8-12 weeks old; conventionally housed with free access to food and water and used according to Macalester College&#39;s IACUC guidelines.&#160;</td>
		</tr>
	</tbody>
</table>

isolation of infiltrating leukocytes from mouse skin using enzymatic digest and gradient separation

Dissociating murine skin into a single cell suspension is essential for downstream cellular analysis such as the characterization of infiltrating immune cells in rodent models of skin inflammation. Here, we describe a protocol for the digestion of mouse skin in a nutrient-rich solution with collagenase D, followed by separation of hematopoietic cells using a discontinuous density gradient. Cells thus obtained can be used for in vitro studies, in vivo transfer, and other downstream cellular and molecular analyses including flow cytometry. This protocol is an effective and economical alternative to expensive mechanical dissociators, specialized separation columns, and harsher trypsin- and dispase-based digestion methods, which may compromise cellular viability or density of surface proteins relevant for phenotypic characterization or cellular function. As shown here in our representative data, this protocol produced highly viable cells, contained specific immune cell subsets, and had no effect on integrity of common surface marker proteins used in flow cytometric analysis.

콜라게나 D 미디어 처리 대조군과 비교시 비장 세포는 T 세포상의 CD4 및 CD8 α 비슷한 수준의 치료를 보여 먼저, T 세포의 서브 세트에 리니지 및 활성화 마커의 주파수 및 풍부 표면에 콜라게나 D의 잠재적 효과는 대조군으로 차 림프 조직을 사용하여 평가 하였다. ?...

Watch this Scientific Journal Video about Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation at JoVE.com

Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation

효소 다이제스트 및 그라데이션 분리를 사용하여 마우스 피부에서 침투 백혈구의 분리

This protocol describes enzymatic digestion of mouse skin in nutrient-rich medium followed by gradient separation to isolate leukocytes. Cells thus derived can be used for diverse downstream applications. This is an effective, economical, and improved alternative to tissue dissociation machines and harsher trypsin and dispase-based tissue digestion protocols.

Dissociating murine skin into a single cell suspension is essential for downstream cellular analysis such as the characterization of infiltrating immune ...

The overall goal of this procedure is to isolate skin-infiltrating leukocytes using enzymatic digestion and density gradient separation. This method can be applied to the investigation of cellular and molecular mechanisms in many mouse models of skin pathologies, including atopic dermatitis. Though this method can provide insight into skin-infiltrating leukocyte biology, it can also be applied to the investigation of other resident cell populations.Generally, individuals new to this method will struggle with the skin harvesting, which includes a complete removal of the excess adipose and connective tissues. Visual demonstration of this method is critical, as skin harvesting and density gradient handling are difficult to learn, and require precision and practice. At the appropriate time point after treatment, use 10 centimeter surgical scissors to carefully excise a 25 by 25 millimeter area of shaved flank skin from an adult eight to twelve-week-old female ND4 Swiss-Webster mouse.Use a clean, sharp surgical blade to scrape away the excess fat and connective tissue from the skin, then transfer the skin into a 50 milliliter conical tube containing 10 milliliters of room temperature HBSS supplemented with EDTA, HEPES, and FBS, and predigest the tissue on a rotating plate for 30 minutes in a dry, non-gassed, 37 degree Celsius incubator. At the end of the incubation, vortex the tube vigorously for ten seconds and filter the tissue slurry through a 70 micron strainer. Next, use a pair of forceps to transfer the flank skin pieces from the strainer into a new 50 milliliter tube containing ten milliliters of fresh, room temperature HBSS media, supplemented with EDTA, HEPES, and FBS, then use a 12.5 centimeter pair of surgical dissecting scissors to mince each piece of flank skin so that the average piece of tissue is approximately 2.2 by 2.2 millimeters in size.When all of the pieces have been minced, return the tube to the rotating plate for another 30 minute incubation. Vortex the tube vigorously for 10 seconds at the end of the incubation and filter the contents through the 70 micron strainer. Collect the remaining flank skin pieces in a new 50 milliliter tube containing 10 milliliters of HBSS supplemented with 0.7 milligrams per milliliter of Collagenase D, and return the tube to the rotating plate for another 30 minute incubation.After another 10 seconds of vortexing, filter the contents of the tube again, this time discarding the debris on the strainer, then fill the tube with up to 50 milliliters of HBSS, EDTA, HEPES, FBS, spin down the cells and decant the supernatant. Add three milliliters of room temperature, 67 percent density gradient centrifugation medium in PBS to a new 15 milliliter conical tube, and resuspend the cell pellet in five milliliters of room temperature 44 percent density gradient centrifugation medium in HBSS with phenol red. Setting the pipettor to the lowest speed, place the pipette on the side of the tube and gently layer the cells on top of the 67 percent density gradient centrifugation medium.When working with density gradients, take care to avoid making bubbles and to minimize any disruptions to the gradient interface. Then separate the cells, using a five milliliter transfer pipette to collect the cells at the interface at the end of the centrifugation. As gently layering the gradient and carefully extracting the interface of the gradients are critical steps, it is wise to practice the density gradient centrifugation medium layering and interface removal with the suspension of murine splenocytes before attempting the procedure with the experimental cells of interest.Finally, wash the collected cells in ice-cold PBS supplemented with FBS in a 15 milliliter conical tube and count the number of viable cells by trypan blue exclusion. CD4 and CD8-alpha surface expression are not affected by Collagenase D digestion, as the same number of CD4 positive and CD8-alpha positive cells is detected with or without enzymatic digestion. Further, Collagenase D digestion exhibits no effect on the surface density of the T cell activation markers CD44 and CD62L on CD4 positive T cells.After gradient separation, about 250 viable immune cells per square millimeter of flank tissue are isolated from Oxazolone, or Ox-challenged mice, versus only four viable immune cells per square millimeter of tissue in vehicle-challenged mice, previously sensitized with Ox.95 percent of the low forward and side scatter single cells in Ox-challenged mice, and 71 percent of these cells in vehicle-challenged controls are live CD45 positive cells, equivalent to an approximately 67 fold increase in the infiltrating CD45 positive immune cell population in the flank skin following three daily Ox challenges. Indeed, Ox challenge results in the pronounced infiltration of T cells and neutrophils into the flank skin, with 42 percent of the single, live CD45 positive cells in Ox-treated animals, apparent Gr-1 positive neutrophils, and approximately 84 percent CD3 positive T cells expressing CD8-alpha, or CD4, compared to control, ethanol-treated animals. Once mastered, this technique can be completed in about three hours if it's performed properly.While attempting this procedure, it's important to remember to handle the density gradient carefully. Don't forget to always use room temperature density gradient centrifugation medium when performing this procedure. Following this procedure, other methods like flow cytometric analysis, cell sorting, cell transfer, or in vitro cell culture can be performed to evaluate the phenotype and function of the skin-infiltrating leukocytes.After watching this video, you should have a good understanding of how to isolate skin-infiltrating leukocytes using enzymatic digestion and density gradient centrifugation.

Research

JoVE Journal

Immunology and Infection

Isolation of Brain-infiltrating Leukocytes

브레인 - 침투 Leukocytes의 분리

We describe a method for preparing brain infiltrating leukocytes (BILs) from mice. We demonstrate how to infect mice with Theiler's murine ...

연구

면역학 및 감염병학

Isolation of Lymphocytes from Mouse Genital Tract Mucosa

마우스 성기 요로 점막에서 림프구의 분리

Mucosal surfaces, including in the gastrointestinal, urogenital, and respiratory tracts, provide portals of entry for pathogens, such as viruses and ...

Isolation and Analysis of Brain-sequestered Leukocytes from Plasmodium berghei ANKA-infected Mice

Isolation and Analysis of Brain-sequestered Leukocytes from Plasmodium berghei ANKA-infected Mice

에서 뇌 - 분리 된 백혈구의 분리 및 분석<em&gt; Plasmodium berghei</em&gt; ANKA에 감염된 마우스

We describe a method for isolation and  characterization of adherent inflammatory cells from brain blood vessels of P. berghei ANKA-infected mice. ...

Bioenergetics and the Oxidative Burst: Protocols for the Isolation and Evaluation of Human Leukocytes and Platelets

인간 백혈구 및 혈소판의 분리 및 평가를위한 프로토콜 : 생물 에너지와 산화 버스트

Mitochondrial dysfunction is known to play a significant role in a number of pathological conditions such as atherosclerosis, diabetes, septic shock, and ...

Analyzing the Effects of Stromal Cells on the Recruitment of Leukocytes from Flow

흐름에서 백혈구의 모집에 기질 세포의 영향 분석

Stromal cells regulate the recruitment of circulating leukocytes during inflammation through cross-talk with neighboring endothelial cells. Here we ...

Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture

Lymphocyte Isolation from Human Skin for Phenotypic Analysis and Ex Vivo Cell Culture

표현형 분석 및 대한 인간의 피부에서 림프구 분리<em&gt; 전의 VIVO</em&gt; 세포 배양

Human skin has an important barrier function and contains various immune cells that contribute to tissue homeostasis and protection from pathogens. As the ...

Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions

다른 마우스 피부 지역에서 Cytofluorimetric 분석을위한 단일 세포 현탁액의 준비

The skin is a barrier organ that interacts with the external environment. Being continuously exposed to potential microbial invasion, the dermis and ...

Isolation of Leukocytes from the Human Maternal-fetal Interface

인간의 산모 - 태아 인터페이스에서 백혈구의 분리

Pregnancy is characterized by the infiltration of leukocytes in the reproductive tissues and at the maternal-fetal interface (decidua basalis and decidua ...

Isolation of Leukocytes from the Murine Tissues at the Maternal-Fetal Interface

산모 - 태아 인터페이스에서 쥐의 조직에서 백혈구의 분리

Immune tolerance in pregnancy requires that the immune system of the mother undergoes distinctive changes in order to accept and nurture the developing ...

Isolation and Flow Cytometric Analysis of Glioma-infiltrating Peripheral Blood Mononuclear Cells

분리 및 신경 교종-침투 말초 혈액 단핵 세포의 유세포 분석

Our laboratory has recently demonstrated that natural killer (NK) cells are capable of eradicating orthotopically implanted mouse GL26 and rat CNS-1 ...