?實(shí)驗(yàn)小鼠是進(jìn)行科學(xué)研究常用的實(shí)驗(yàn)動(dòng)物,小鼠不僅和人的基因具有相似度初步建立,而且具有體型小便利性、易飼養(yǎng)建設項目、生長(zhǎng)繁殖快統籌推進、操作管理方便的優(yōu)點(diǎn)。隨著小鼠在科學(xué)研究中的廣泛應(yīng)用技術創新,科研人員培育出了許多小鼠品種及品系深入交流研討。在進(jìn)行研究計(jì)劃時(shí),選擇合適品系的小鼠建立動(dòng)物疾病模型廣泛應用,是實(shí)驗(yàn)設(shè)計(jì)的關(guān)鍵基礎(chǔ)一步關註度。
BALB/c小鼠?:
?遺傳背景?:1913年美國(guó)國(guó)立腫瘤研究所培育,經(jīng)過(guò)多代近交繁殖哪些領域。
?特征?/用途?:?
1.性格溫順敢於挑戰,易于繁殖,雌雄體重差異薪⒑屯晟?√峁┝俗裱?;
2.乳腺腫瘤發(fā)病率低(3%),當(dāng)用乳腺腫瘤病毒(MTV)誘導(dǎo)時(shí)發(fā)病率將增高產業。對(duì)礦物油誘導(dǎo)漿細(xì)胞瘤敏感滿意度,廣泛應(yīng)用于雜交瘤和單克隆抗體的生產(chǎn);
3.易患慢性肺炎可持續;
4.有自發(fā)高血壓癥,老年鼠心臟病變,雌雄鼠均有動(dòng)脈硬化構建,幾乎全部20月齡的雄鼠脾臟均有淀粉樣變創新科技;
5.對(duì)放射線敏感,應(yīng)用于核醫(yī)學(xué)共創輝煌。
?C57BL/6小鼠?:
?遺傳背景?:1921年C.C.Little用Abby Lathrop小鼠培育而成具有重要意義。
?特征?/用途?:
1.已經(jīng)完成基因組測(cè)序的小鼠品系;
2.品系穩(wěn)定大部分、容易繁殖強大的功能;
3.試驗(yàn)結(jié)果精度高,可比性好解決方案,應(yīng)激反應(yīng)均一優勢;
4.構(gòu)建基因修飾動(dòng)物模型,可保證遺傳背景的高度穩(wěn)定性和實(shí)驗(yàn)數(shù)據(jù)一致性增產。
有了這些基礎(chǔ)知識(shí)背景便利性,我們?cè)跇?gòu)建模型時(shí),設(shè)計(jì)實(shí)驗(yàn)時(shí)高產,參考文獻(xiàn)時(shí)信息化技術,評(píng)價(jià)數(shù)據(jù)時(shí),就能更準(zhǔn)確良好。尤其我們使用荷蘭Liposoma巨噬細(xì)胞清除劑Clodronate Liposomes氯膦酸鹽脂質(zhì)體清除巨噬細(xì)胞時(shí)逐步顯現,我們除了關(guān)注這些因素,還需要考慮小鼠的周齡引領。巨噬細(xì)胞的高度異質(zhì)性和可塑性傳遞,也覺(jué)得巨噬細(xì)胞的多功能性和多面性,動(dòng)態(tài)性和適應(yīng)性勞動精神。比如巨噬細(xì)胞可以質(zhì)控造血干細(xì)胞(見(jiàn)Science文獻(xiàn))開展攻關合作,巨噬細(xì)胞可以和神經(jīng)元一樣調(diào)控運(yùn)動(dòng)(見(jiàn)Natuer文獻(xiàn))。在這些頂刊的重大研究發(fā)現(xiàn)中預下達,研究人員都使用荷蘭Liposoma的巨噬細(xì)胞清除劑(CP-005-005)貨號(hào)的有效手段。此外,巨噬細(xì)胞等免疫細(xì)胞方案,在不同品系關鍵技術,不同周齡,不同性別深入,也是差異很大技術研究。見(jiàn)如下這篇文獻(xiàn)的基礎(chǔ)研究。
小鼠選擇
品系:C57BL/6和BALB/c
性別:雄性鼠
周/月齡:1開展研究、3姿勢、5相互融合、10和18月齡
處死方式:通過(guò)CO2窒息處死
樣品:外周血和脾臟
流式細(xì)胞術(shù)分析
1、樣本制備:
外周血單個(gè)核細(xì)胞(PBMC):血液PBS稀釋后,使用ACK裂解液去除紅細(xì)胞•脾細(xì)胞:機(jī)械分離后同樣使用ACK裂解液處理
2綠色化、抗體標(biāo)記: 使用熒光標(biāo)記的單克隆抗體組合:
FITC標(biāo)記: CD4不同需求、CD19、MHC II保持穩定、CD11b
PE標(biāo)記: CD3總之、CD8、CD11c支撐作用、CD44
PerCP標(biāo)記: CD3研學體驗、CD19
研究結(jié)果
參考文獻(xiàn):
Pinchuk LM, Filipov NM. Differential effects of age on circulating and splenic leukocyte populations in C57BL/6 and BALB/c male mice. Immun Ageing. 2008 Feb 11;5:1. doi: 10.1186/1742-4933-5-1. PMID: 18267021; PMCID: PMC2268915.
Using a multipoint age comparison approach, cells from the two major immune system compartments, peripheral blood and spleen, and flow cytometry analysis, we found several principal differences in T cell and professional antigen presenting cell (APC) populations originating from a prototypical T helper (Th) 1 mouse strain, C57BL/6, and a prototypical Th2 strain, BALB/c. For example, regardless of age, there were strain differences in both peripheral blood mononuclear cells (PBMC) and spleens in the proportion of CD4+ (higher in the BALB/c strain), CD8+ T cells and CD11b+/CD11c+ APC (greater in C57BL/6 mice). Other differences were present only in PBMC (MHC class II + and CD19+ were greater in C57BL/6 mice) or differences were evident in the spleens but not in circulation (CD3+ T cells were greater in C57BL/6 mice). There were populations of cells that increased with age in PBMC and spleens of both strains (MHC class II+), decreased in the periphery and spleens of both strains (CD11b+) or did not change in the PBMC and spleens of both strains (CD8+). We also found strain and age differences in the distribution of na?ve and memory/activated splenic T cells, e.g., BALB/c mice had more memory/activated and less naive CD8+ and CD4+ T cells and the C57BL/6 mice.
Our data provide important information on the principal differences, within the context of age, in T cell and professional APC populations between the prototypical Th1 mouse strain C57BL/6 and the prototypical Th2 strain BALB/c. Although the age-related changes that occur may be rather subtle, they may be very relevant in conditions of disease and stress. Importantly, our data indicate that age and strain should be considered in concert in the selection of appropriate mouse models for immunological research.
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