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Overview of Cell Proliferation

Cell proliferation is an increase in cell number due to cell division, or cytokinesis, the final step of the cell cycle. Cell proliferation is necessary for normal tissue development and maintenance over the lifespan.

Cell proliferation is a tightly regulated process, with many different proteins controlling cell cycle checkpoints. Genetic mutations found in cancer cells cause uncontrolled cellular proliferation.

如何测量细胞增殖

The cell cycle is composed of multiple phases, including the G1 phase where cells increase in size, the S phase where DNA is newly synthesized, the G2 phase where further cell growth occurs, and finally the M, or mitosis, phase where the cell ultimately divides. Some cells exit the cell cycle and stop dividing altogether; this is known as quiescence or resting state (G0)

细胞周期各阶段

Phases of the Cell Cycle: G1 - 大小增加;S - DNA 合成和复制;G2 - 生长;M - 有丝分裂和细胞分裂;G0 - 休眠期。

Two of the most common ways of measuring cell proliferation include cell cycle assays and proliferation assays, which rely on the incorporation of propidium propidium iodide or BrdU into DNA:

Assay What is Measured

Cell Cycle Assay Dyes

Propidium Iodide

7-AAD

Quantifies the number of cells at different stages of the cell cycle based on the profile of their DNA.
Propidium Iodide (PI)/RNase Staining Solution #4087

Propidium Iodide (PI)/RNase Staining Solution #4087: 使用 Phospho-Histone H3 (Ser10) Antibody (Alexa Fluor® 488 Conjugate) #9708 和 Propidium Iodide (PI)/RNase Staining Solution 对未经处理的 Jurkat 细胞进行流式细胞分析(DNA 含量)。

BrdU Cell Proliferation Assay Kit

Measures the level of DNA synthesis occuring during cellular proliferation
BrdU Cell Proliferation Assay Kit #6813:

BrdU Cell Proliferation Assay Kit #6813: C2C12 细胞按不同密度接种在 96 孔板的无血清培养基中,并孵育过夜。按不同浓度往平板上添加血清,并孵育细胞 24 小时。最后,往平板中加入 10 μM BrdU,并孵育细胞 4 小时。

Markers to Determine Cell Proliferation

Another common method for measuring cell proliferation is by examining the expression levels of cell cycle proteins and other proteins required for proliferation. Some common protein markers for proliferating cells include:

Proliferation Marker Proliferation Marker Description
Proliferating cell nuclear antigen (PNCA) DNA sliding clamp protein that plays an important role in DNA replication
PCNA (D3H8P) XP<sup>®</sup> Rabbit mAb #13110

PCNA (D3H8P) XP® Rabbit mAb #1311: 使用 PCNA (D3H8P) XP® Rabbit mAb (绿色)和 β-Catenin (L54E2) Mouse mAb (IF Preferred) #2677(红色)对显示肠隐窝中增殖细胞染色的大鼠小肠细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。

Ki-67 Non-nuclear histone protein only expressed at high levels in proliferating cells
Ki-67 (D2H10) Rabbit mAb (IHC Specific) #9027

Ki-67 (D2H10) Rabbit mAb (IHC Specific) #9027: 使用 Ki-67 (D2H10) Rabbit mAb (IHC Specific) 对石蜡包埋的人结肠癌进行免疫组织化学分析。

Phospho-histone H3 Nuclear histone protein that is phosphorylated in proliferating cells
Phospho-Histone H3 (Ser10) (D7N8E) XP<sup>®</sup> Rabbit mAb #53348

Phospho-Histone H3 (Ser10) (D7N8E) XP® Rabbit mAb #53348: 使用 Phospho-Histone H3 (Ser10) (D7N8E) XP® Rabbit mAb (上图) 或 Histone H3 (D1H2) Rabbit mAb #4499 (下图),对未经处理的或经 Nocodazole #2190 (100ng/ml,16小时) 处理的 Hela 细胞提取物进行蛋白质印迹分析。该抗体的磷酸特异性的显示是进一步使用 λ-磷酸酶处理细胞裂解物。

β-galactosidase activity β-galactosidase activity is associated with senescent cells that are not proliferating
Senescence β-Galactosidase Staining Kit #9860:

Senescence β-Galactosidase Staining Kit #9860: β-Galactosidase staining at pH 6 on senescent WI38 cells at population doubling 36 (right).

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