台灣轉錄科技有限公司    

BD Vacutainer CPT cell separation 

BD Vacutainer® CPT™ Cell Preparation Tube with Sodium Citrate

REF 362753   8 mL Draw Capacity  with Sodium heparin (16 x 125mm tube Size)  60sets/case

REF 362760   4 mL Draw Capacity with Sodium Citrate  (13 x 100mm tube Size)  60sets/case

REF 362761   8 mL Draw Capacity  with Sodium Citrate (16 x 125mm tube Size) Steriled  60sets/case

科学研究案例

·         主题:自体细胞生长因子提取 Platelet Rich Plasma (PRP)

·         实验目的：主要目的在于提取血小板中的生长因子(Growth Factor)：IGF1、IGF2、EGF、VEGF、TGF-alpha、TGF-beta、PDGF-aa、PDGF-bb、PDGF-ab等。

·         原理：是将自己身体内静脉血管抽取出全血，利用CPT比重梯度试管，将含有高浓度血小板的全血，经过304XG离心取得个别血球比重分层，再经由离心与纯化之后，取出高浓度的血小板，也就是俗称PRP自体生长因子(Platelet Rich Plasma)释放出生长因子。 

·         实验方法：

• Collect blood into the Tube using the standard technique for BD Vacutainer® Brand Blood Collection Tubes (see Venipuncture Technique & Sample Collection section and Prevention of Backflow section).After collection, store Tube upright at room temperature until centrifugation. Blood samples should be centrifuged within two hours of blood collection for best results.Centrifuge Tube/blood sample at room temperature (18-25° C) in a horizontal rotor (swing-out head) for a minimum of 20 minutes at 1500 to 1800 RCF (Relative Centrifugal Force).

• After collection, store Tube upright at room temperature until centrifugation. Blood samples should be centrifuged within two hours of blood collection for best results.

• Centrifuge Tube/blood sample at room temperature (18-25° C) in a horizontal rotor (swing-out head) for a minimum of 20 minutes at 1500 to 1800 RCF (Relative Centrifugal Force).

• After centrifugation, mononuclear cells and platelets will be in a whitish layer just under the plasma layer (see above figure). Aspirate approximately half of the plasma without disturbing the cell layer. Collect cell layer with a Pasteur pipette and transfer to a 15 mL size conical centrifuge Tube with cap. Collection of cells immediately following centrifugation will yield best results.

• An alternative procedure for recovering the separated mononuclear cells is to resuspend the cells into the plasma by inverting the unopened BD Vacutainer® CPT™ Tube gently 5 to 10 times. This is the preferred method for storing or transporting the separated sample for up to 24 hours after centrifugation. To collect the cells, open the BD Vacutainer® CPT™ Tube and pipette the entire contents of the Tube above the gel into a separate vessel.

• Keyword：自体细胞、PRP (Platelet Rich Plasma)、干细胞、生长因子、CPT tube、血小板、干细胞治疗、组织修复、自体胶原蛋白、PRP美容、PRP除皱。

Suggested  Cell   Washing   Steps       :

1. Add PBS to bring volume to 15 mL. Cap Tube. Mix cells by inverting Tube 5 times.

2. Centrifuge for 15 minutes at 300 RCF. Aspirate as much supernatant as possible without disturbing cell pellet.

3. Resuspend cell pellet by gently vortexing or tapping Tube with index finger.

4. Add PBS to bring volume to 10 mL. Cap Tube. Mix cells by inverting Tube 5 times.

5. Centrifuge for 10 minutes at 300 RCF. Aspirate as much supernatant as possible without disturbing cell pellet. Resuspend cell pellet in the desired medium for subsequent procedure.

·         实验结果：经过流式细胞测定仪 (FlowCytometry) 对各细胞功能测试，测出高纯度单核球、CD61结合测定高浓度血小板(78%)，经过干细胞结合，生长因子促进及浓缩等作用，加速组织再生，伤口愈合与重建；将取得良好效果。

·         讨论：本项研究对血小板浓缩，血小板提取，血小板分离，利用比重梯度法经过离心清洗可达到良好效果，血小板生长因子促进干细胞分化，对组织细胞再生也取得良好效果；对日后再进行同血小板功能研究及医学美容科学研究，取得跨越性进展基础。