• Genetic origin: The pgsA-745 cell line was created by treating wild-type CHO-K1 cells with a mutagen (ethylmethanesulfonate) and then screening for mutants with defects in proteoglycan synthesis.

• Xylosyltransferase deficiency: The primary defect is a deficiency in xylosyltransferase (XylT), an enzyme essential for adding the first sugar residue in the synthesis of GAG chains, including heparan sulfate (HS) and chondroitin sulfate (CS). Due to this, the cells are unable to produce GAGs.

• Secondary mutation: More recent whole-genome sequencing has revealed that pgsA-745 cells also carry a secondary mutation: a deletion in the gene for laminin subunit $\alpha$2 (Lama2). This deletion affects the cell's ability to be invaded by certain bacteria, a trait initially attributed only to the GAG deficiency.

• Female origin: The cell line was derived from the ovary of a female Chinese hamster.

• Spontaneously immortalized: Like its parent line, pgsA-745 is an immortalized cell line.

The pgsA-745 cell line is widely used in biomedical research to study the functions of GAGs and proteoglycans. Some applications include:

• Cell adhesion and signaling: Studying the role of proteoglycans in cell-extracellular matrix interactions.

• Pathogen invasion studies: Investigating how the absence of GAGs affects cell invasion by various pathogens, including bacteria and viruses.

• Recombinant protein production: Used as a host cell line, particularly for producing proteins that do not require GAGs for proper function. The line is also studied for its resistance to infection.

• Glycosaminoglycan biology: Characterizing the specific roles of different GAGs in various cellular processes by comparing the mutant cells with wild-type CHO-K1 cells.

• Biosafety level: 1.

• Cryostorage: For long-term storage, frozen vials must be stored in the vapor phase of liquid nitrogen to preserve viability.

• Resuscitation: Upon thawing, the cryoprotective agent should be removed promptly via a quick spin and the cells resuspended in the appropriate complete growth medium.

• Medium and atmosphere: A complete medium, such as Ham's F12 with 10% fetal bovine serum, is used, along with a humidified 5% CO2 atmosphere.

BioVector NTCC質(zhì)粒載體菌種細(xì)胞蛋白抗體基因保藏中心

BioVector NTCC Inc.

TEL: 400-800-2947, 189-0126-8599

E-mail: [email protected]

http://www.nedfriskphoto.com

• 遺傳起源：pgsA-745 細(xì)胞系通過用誘變劑（乙基甲磺酸）處理野生型 CHO-K1 細(xì)胞，然后篩選蛋白聚糖合成缺陷的突變體而創(chuàng)建。

• 木糖基轉(zhuǎn)移酶缺陷：主要缺陷是木糖基轉(zhuǎn)移酶（XylT）的缺乏。該酶是合成 GAG 鏈（包括肝素硫酸和硫酸軟骨素）中添加第一個糖殘基所必需的。因此，該細(xì)胞無法產(chǎn)生 GAGs。

• 次要突變：最近的全基因組測序顯示，pgsA-745 細(xì)胞還攜帶一個次要突變：層粘連蛋白亞基 $\alpha$2（Lama2）基因的缺失。這一缺失會影響細(xì)胞被某些細(xì)菌侵入的能力，這一特征最初只被歸因于 GAG 缺陷。

• 雌性起源：該細(xì)胞系源自雌性中國倉鼠的卵巢。

• 自發(fā)永生化：與其親本細(xì)胞系一樣，pgsA-745 是一種永生化細(xì)胞系。

• 細(xì)胞黏附和信號傳導(dǎo)：研究蛋白聚糖在細(xì)胞-細(xì)胞外基質(zhì)相互作用中的作用。

• 病原體入侵研究：研究 GAGs 的缺失如何影響各種病原體（包括細(xì)菌和病毒）對細(xì)胞的入侵。

• 重組蛋白生產(chǎn)：用作宿主細(xì)胞系，特別是用于生產(chǎn)不需要 GAGs 發(fā)揮正常功能的蛋白。該細(xì)胞系也因其抗感染性而受到研究。

• 糖胺聚糖生物學(xué)：通過比較突變細(xì)胞和野生型 CHO-K1 細(xì)胞，表征不同 GAGs 在各種細(xì)胞過程中的具體作用。

• 生物安全級別：1 級。

• 凍存：為了長期保存，冷凍管必須儲存在液氮氣相中，以保持細(xì)胞活力。

• 復(fù)蘇：解凍后，應(yīng)通過快速離心立即去除冷凍保護(hù)劑，并將細(xì)胞重新懸浮在適當(dāng)?shù)耐耆L培養(yǎng)基中。

• 培養(yǎng)基和培養(yǎng)環(huán)境：使用含 10% 胎牛血清的 Ham's F12 等完全培養(yǎng)基，以及 5% CO2 的濕潤氣氛。

BioVector NTCC質(zhì)粒載體菌種細(xì)胞蛋白抗體基因保藏中心

BioVector NTCC Inc.

TEL: 400-800-2947, 189-0126-8599

E-mail: [email protected]

http://www.nedfriskphoto.com