HyCyte® | 海星生物

发货周期：

干冰冻存管运输3天（1x10⁶cells/冻存管x2）

常温运温1-2周发货 (1x10⁶ cells/T25培养瓶）

该细胞由CollinsSJ从一位患有急性早幼粒细胞性白血病的36岁白人女性的外周血中分离建立；可自发分化，或在丁酸盐、次黄嘌呤、佛波醇肉豆蔻酸(PMA,TPA)、DMSO(1%to1.5%)、放线菌素D和视黄酸的刺激下发生分化；PMA刺激后可分泌TNF-α。该细胞具有吞噬活性和趋化反应，癌基因myc阳性，表达补体受体和FcR。

细胞名称：人原髓细胞白血病细胞(STR鉴定正确)

细胞简称：HL-60

产品货号：TCH-C204

种属来源：人

组织来源：外周血

疾病特征：急性粒-单核细胞白血病

细胞形态：髓母细胞样

生长特性：悬浮生长

培养体系：IMDM+20%FBS+1%P/S

配套培养基货号：TCH-G204

传代比例：1:3-1:6，每2-3天换液一次

传代周期：48-72 h

培养条件：气相：95%空气+5%CO₂，温度：37℃

冻存条件：60%基础培养基+30%FBS+10%DMSO，液氮储存

质量检测：细菌、真菌、支原体检测均为阴性

参考文献：

"Gallagher R, et al. Characterization of the continuous, differentiating myeloid cell line (HL-60) from a patient with acute promyelocytic leukemia. Blood 54: 713-733, 1979. PubMed: 288488

Collins SJ, et al. Terminal differentiation of human promyelocytic leukemia cells induced by dimethyl sulfoxide and other polar compounds. Proc. Natl. Acad. Sci. USA 75: 2458-2462, 1978. PubMed: 276884

Collins SJ, et al. Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture. Nature 270: 347-349, 1977. PubMed: 271272

Aggarwal BB, et al. Human tumor necrosis factor. Production, purification, and characterization. J. Biol. Chem. 260: 2345-2354, 1985. PubMed: 3871770

Nahm MH, et al. Identification of cross-reactive antibodies with low opsonophogocytic activity for Streptoccus pneumoniae. J. Infect. Dis. 176: 698-703, 1997. PubMed: 9291318

Berninghausen O, Leippe M. Necrosis versus apoptosis as the mechanism of target cell death induced by Entamoeba histolytica. Infect. Immun. 65: 3615-3621, 1997. PubMed: 9284127

Aparicio CL, et al. Correction for label leakage in fluorimetric assays of cell adhesion. BioTechniques 23: 1056-1060, 1997. PubMed: 9421636

Mansat V, et al. The protein kinase C activators phorbol esters and phosphatidylserine inhibit neutral aphingomyelinase activation, ceramide generation, and apoptosis triggered by daunorubicin. Cancer Res. 57: 5300-5304, 1997. PubMed: 9393753

Cuthbert JA, Lipsky PE. Regulation of proliferation and Ras localization in transformed cells by products of mevalonate metabolism. Cancer Res. 57: 3498-3504, 1997. PubMed: 9270019

Michael JM, et al. Resistance to radiation-induced apoptosis in Burkitt's lumphoma cells is associated with defective ceramide signaling. Cancer Res. 57: 3600-3605, 1997. PubMed: 9270034

Clark RA, et al. Tenascin supports lymphocyte rolling. J. Cell Biol. 137: 755-765, 1997. PubMed: 9151679

Tiffany HL, et al. Enhanced expression of the eosinophil-derived neurotoin ribonuclease (RNS2) gene requires interaction between the promoter and intron. J. Biol. Chem. 271: 12387-12393, 1996. PubMed: 8647842

Chan YJ, et al. Synergistic interactions between overlapping binding sites for the serum response factor and ELK-1 proteins mediate both basal enhancement and phorbol ester responsiveness of primate cytomegalovirus. J. Virol. 70: 8590-8605, 1996. PubMed: 8970984

Mao M, et al. RIG-E, a human homolog of the murine Ly-6 family, is induced by retinoic acid during the differentiation of acute promyelocytic leukemia cell. Proc. Natl. Acad. Sci. USA 93: 5910-5914, 1996. PubMed: 8650192

Lepley RA, et al. Tyrosine kinase activity modulates catalysis and translocation of cellular 5-lipoxygenase. J. Biol. Chem. 271: 6179-6184, 1996. PubMed: 8626407

Chen H, et al. Octamer binding factors and their coactivator can activate the murine PU.1 (spi-1) promoter. J. Biol. Chem. 271: 15743-15752, 1996. PubMed: 8663022

U.S. Pharmacopeia USP Monographs: Technetium 99mTc Fanolesomab Injection. Rockville, MD: USP32-NF27, 2005"

HL-60细胞培养注意要点

(1) HL-60细胞在高密度下状态比较好，细胞状态好时会吸收一些细胞碎片；

(2) 低密度时细胞状态很差，传代比例要控制，当培养密度达到1x10⁶/mL左右时可传代，细胞状态不好时后面传代可以不用离心；

(3) DMSO会对HL-60刺激分化，建议复苏马上离心去除DMSO；

(4) 培养过程中尽量不动或少动培养瓶；

(5) 血清质量差异可能引起细胞状态变化，建议选用高质量的胎牛血清。

HL-60细胞培养常见问题

1. HL-60严格来讲单个细胞，不成团；

2. 死细胞成团，死细胞释放DNA把细胞黏成团；

3. HL-60细胞生长慢、状态差的解决方法：

(1) 当细胞传代后生长速度慢且状态不佳时，可竖着培养直到培养基变黄，待细胞密度起来后，状态会有所好转。

(2) 若HL-60细胞状态很差，可采用半换液的方式：

以T25瓶子为例，瓶子里有5mL培养基，竖起瓶子静置一段时间（竖瓶前拍拍瓶子，让轻贴底部的细胞团浮起来），待细胞沉底（肉眼观察细胞沉底情况），小心吸出2.5mL的培养基，转移到离心管，离心800rpm 4min，用2.5mL新鲜培养基重悬后再放回原瓶。

收货当天如何处理细胞？

收到细胞后请务必仔细阅读产品资料，了解细胞相关信息，如贴壁特性 (贴壁/悬浮/半贴壁半悬浮)、细胞形态、培养体系、传代比例和换液频率等。