201004659 九、發明說明： 【發明所屬之技術領域】 本發明係有關金液保存的方法，尤其是有關自體血小板乾粉化 保存的方法。 【先前技術】 有關「體外保存血小板」的研究，雖然有相當多的論文發表， 然而因為製程及保存的不易，絕大部分僅止於研發階段尚未有成 ❹功的報告’隨著細胞生物學和分子生物學研究的明朗化，目前已 經證實了由血小板釋出的生長因子在創傷修復過程中具有舉足輕 重的作用’它參與調控創傷修復的各個階段，影響傷口的癒合和 組織的重建，是創傷修復中不可缺少的。 由於jk小板巍含豐富的生長因子，可以誘導並調節細胞的生 長、分化，進而促進骨組織再生與軟組織之癒合，因此醫學臨床 上’常利用血小板濃厚液（Aut〇1〇g〇us伽此⑺删加把pc ) 〇 或富含血小板的血漿(Autolo_s platelet-rich Pl_, PRP) 來促進術後組織的癒合’但為了避免排斥關題，故皆利用來自 於病人本身的血液離心製作成血小板濃厚液，稱為自體血小板濃 厚液(Autologous piatelet c〇ncentrate，Apc)。目前已經有許 多薇商研發出相關的套裝產品，供醫師於手術前先製備病患自體 的PRP，於-定時間内使用完畢，以加速病患組織及傷口的癒合， 但因為PRP的保存方式十分嚴苛，有效期非常的短暫，因此到目 前為止’絕大部分都是於使用前才開始製備，也因為如此，雖然 201004659 知道PRP的醫療功效非凡，但也因為不易保存，只能「現場備製、 立即使用」，而大大地限制了 PRP的市場經營方式及規模。 隨著經濟的起飛及醫療科技的進步，傷口修復的市場日益龐 大，如果能夠克服PRP的短效儲存期，延長PRp的使用週期，這 將是會是一個非常龐大的市場。 有一些血小板保存的方法的專利技術，例如美國專利 5, 736’ 313、台灣專利1270375及台灣公開專利2〇〇7〇〇556等所揭 ❹示者其巾台灣專利1270375揭示的一種血小板體外保護方法， 主要係利用-種簡單而自鋪方法，保護血小板之細胞膜，使存 在於細胞膜表面之特定糖分子受到掩護，使血小板於體外保存一 段時間後活性仍能保持；該方法之富含血小板的血絲源可為蹲 自血液中心的血小板濃驗’或取自人體的全血㈣雜心方法 .分離出之血，〗、板濃厚㈣；此方法可在辦财娜護血小板， 維持其活性’並可在體外做長時期的保存，不會有化學殘餘沈積 ❹物質’故減雜處理過的血小板，沒摘產物絲的顧慮，且 活性保持完整’可作為體外研究其細胞特性的題材。 另台灣公開專利200700556揭示的體外保存血小板、紅血球、 無核細胞與含A小板組成物之反應劑和方法，係提供一長期保存 血j板之方法’其步驟包括(a)將抗凝血劑、冷凝沈澱物以及凝血 酶加入-般生理食鹽水中形成—溶液；⑹將—含有血小板之介質 加入步驟(a)之溶液並置形成—含有血小板之溶液；以及⑹ 201004659 凍乾步驟(b)所形成之含有血小板溶液。 上述兩專利技術雖分別揭示液體狀血小板的保存方法及乾粉 狀血小板的保存方法’但其使用的血小板係取自血液中心等多數 個捐血者混合的血小板，並非針對某一特定個人自體的血小板加 以保存，因此無法符合個人自體血小板保存的需求。 【發明内容】 為了針對個人自體的血小板加以保存，以備將來供自已使 ❹ 用，而提出本發明。 本發明的主要目的，在提供一種自體血小板乾粉化保存的方 法’使個人可方便的保存自體的血小板。 本發明的另一目的，在提供一種自體血小板乾粉化保存的方 法’使個人可方便進—步隨時彻自體乾粉化的到、板改善自已 的身體。 本發明的又一目的，在提供一種自體血小板乾粉化保存的方 β 法’使個人抽錢可進—步保細餘的血小板。 本發明的其他目的、功效，請參關式及實細，詳細說明 如下。 【實施方式】 Μ參閱圖丨所示^本發明自體血小板乾粉化保存的方法，主 要係使一般的個人(活體)能夠保存自體的血小板者，包括如下步 驟： (a)以分離式捐血機抽取單-频的血液； 201004659 取㈣ι物概鼓岭㈣触血液分離 出血小板加以另外儲存，而將紅血球及血漿送回活體； (C)使待檢查之血液進行血液病毒檢查、醫院例行之血液檢查 (Routine bl(xxl tests)等各種需要的錢檢查，並使儲存的 •到、錄歧聽馳岐小板，贿護血她，維持其活性； ⑷判疋補檢查之血㈣錢縣檢查是否合格，若血液病毒檢 查不合格’則終止血小板的保存作業⑷；若血液病毒檢查合 ❹ 格，則進入步驟（f)，· ⑴使液體保護狀的血小板乾粉化，形成自體乾粉化企小板，以 供長期保存。 上述麵（a)⑽式捐血難取的錢個於製備該活體的 整型手術使用的血小板漠厚液或富含金小板的血聚。而步驟⑷ 中製成自體乾粉化的血小板，係利用該活體的整型手術使用後剩 餘的且經血液病毒檢查合格的血小板濃厚液或富含血小板的血漿 ❹其中之一的血小板所製成者。 上述步驟（b)中的液體保護狀的血小板可利用前述台灣專利 1270375所揭示的技術製成者，例如包括下列步驟： • (1)先將5-30ml抗凝劑加入100ml生理食鹽水（N〇rmal sanne) 中’形成含有5-30%抗凝劑的溶液，搖晃後靜置五分鐘； (2)加入 〇. 〇〇卜5ml 冷凍沈殿劑(Cryoprecipitate ; Cryo， fibrinogen)於上述溶液内，搖晃後靜置一分鐘； 8 201004659 (3) 加入0. 〇(H-5ml的凝血酶(Thrombin)，搖晃後靜置一分鐘； (4) 取血小板彡辰尽液或畐含血小板的血漿，放入上述溶液内，靜置 十分鐘； (5) 置於4°C下或-20°C下保存。 •或包括下列步驟： (1)先將20ml抗凝劑加入100ml生理食鹽水中，形成含有2〇%抗 凝劑的溶液，搖晃後靜置五分鐘； 〇 (2)加入〇·〇〇lml冷凍沈澱劑於上述溶液内，搖晃後靜置一分鐘； (3) 加入5ml的凝血酶，搖晃後靜置一分鐘； (4) 取血小板濃厚液或富含血小板的血漿，放入上述溶液内，靜置 十分鐘； (5) 置於-20°C下保存。 或包括下列步驟： (1) 先將10ml抗凝劑加入l〇〇mi生理食鹽水中，形成含有1〇%抗 ❹ 凝劑的溶液，搖晃後靜置五分鐘； (2) 加入3ml冷凍沈澱劑於上述溶液内，搖晃後靜置一分鐘； (3) 加入0· 01的凝血酶，搖晃後靜置一分鐘； .(4)取血小板濃厚液或富含血小板的血漿，放入上述溶液内，靜置 十分鐘； (5)可置於4它下保存。 或包括下列步驟： 201004659 (1) 先將5ml抗凝劑加入l〇〇mi生理食鹽水中，形成含有5%抗凝 劑的溶液，搖晃後靜置五分鐘； (2) 加入3ml冷凍沈澱劑於上述溶液内，搖晃後靜置一分鐘； (3) 加入0.0001的凝血酶，搖晃後靜置一分鐘； (4) 取血小板濃厚液或富含金小板的血聚，放入上述溶液内，靜置 十分鐘； (5) 可置於4°C下保存。 Ο 上述各步驟（1)中所使用之抗凝劑可採用抗血液凝固擰檬酸 鹽葡萄糖液(Acid Citrate Dextrose, ACD-A)或檸檬酸鱗酸葡萄 糖(Citrate Phosphate Dextrose, CPD)。 上述步驟（f)中的自體乾粉化的血小板可利用前述台灣公 開專利200700556所揭示的技術製成者，例如包括下列步驟： (1)先將20ml之ACD—A加入100ml之生理食鹽水中，形成含有2〇% 抗凝劑之溶液，搖晃後靜置五分鐘； ® (2)加入〇. 〇〇lml冷凝沈澱物於前述抗凝劑溶液中，搖晃後靜置五 分鐘； (3) 加入5ml凝血酶，搖晃後靜置五分鐘； (4) 將血小板濃厚液或富含血小板的血漿加入步驟(3)之溶液，靜 置五分鐘； (5) 最後將步驟(4)之溶液凍乾成粉末後保存。 本發明自體血小板乾粉化保存的方法，可達成下列數種功效： 201004659 1·方便保存自體的灰小板 ^發明可枝個场妓贿自魏粉倾_ 2·減少PRP的浪費與污染 存以備將來改善自⑽聽之用，魏存_可達二年以上聽 ^上爾刪現場備製，且—次収，若當次未用完即需 廢棄，造成PRP的浪費與污染。利用本發明可將未用完的 PRP製 Ο 、粉劑生長因子後保存，完全不會造成pRp的浪費與污染。 3.降低使用者採灰的頻率 使用者每次需要PRP日夺皆要進行採血，採蝴率麵。利用本發 月可隨時提供使用者已保存之自體血小板，無需每次需要時， 都進行採血，可降低採血機率。 4.操作簡易 每-人需要PRP時，皆需進行採企、離心製作的流程，耗時約需3〇 λ鐘。利用本發明製作之乾粉企小板，復水還原後即可使用，操 ❹作相當簡易。 5.節省費用 .以傳統方法從全金中收集PRP，只能使用一次，每次的成本需要 7500元’若以本發明高用量的方式進行處理，每次以正常用量估 算’可使用100次以上，對於2或3個月需要進行一次的醫學美 容治療手術的使用者而言’只要進行一次自體乾粉化的血小板的 製作’即可使用1〇〇次以上的術後保養，其節省費用效益相當明 201004659 顯。 以上所記載，僅為利用本發明技術内容之實施例，任何熟悉 本項技藝者運用本發明所為之修飾、變化，皆屬本發明主張之專 利範圍，而不限於實施例所揭示者。 201004659 【圖式簡單說明】 圖1為實施本發明自體血小板乾粉化保存的方法的流程圖。 【主要元件符號說明】 (a)、（b)、（c)、（d)、（e)、（f )為實施本發明的各步驟編號201004659 IX. Description of the invention: [Technical field to which the invention pertains] The present invention relates to a method for preserving gold liquid, and more particularly to a method for dry powder preservation of autologous platelets. [Prior Art] Although there are quite a few papers on the study of "preserved platelets in vitro", because the process and preservation are not easy, most of them only report on the development stage. And the clarification of molecular biology research, it has been confirmed that the growth factor released by platelets plays a pivotal role in the wound repair process. It is involved in the regulation of various stages of wound repair, affecting wound healing and tissue reconstruction, is trauma Indispensable in the repair. Because jk small plate is rich in growth factors, it can induce and regulate cell growth and differentiation, and then promote bone tissue regeneration and soft tissue healing. Therefore, medical use of platelet thick liquid (Aut〇1〇g〇us gamma) (7) add PC) or platelet-rich plasma (Autolo_s platelet-rich Pl_, PRP) to promote postoperative tissue healing 'but in order to avoid exclusion, it is made using the blood from the patient itself. The platelet thick solution is called Autologous piatelet c〇ncentrate (Apc). At present, many Weishang have developed related kits for doctors to prepare the patient's own PRP before surgery, and use it in a fixed period of time to accelerate the healing of the patient's tissues and wounds, but because of the preservation of PRP. The method is very strict and the validity period is very short. So far, most of them are prepared before use. Because of this, although 201004659 knows that PRP has extraordinary medical effects, it is also difficult to save. Prepared and used immediately, which greatly limits the market operation mode and scale of PRP. With the economic take-off and advances in medical technology, the market for wound repair is growing. If it can overcome the short-term storage period of PRP and extend the life cycle of PRp, it will be a very large market. There are some patented techniques for the method of platelet storage, such as U.S. Patent No. 5,736,313, Taiwan Patent No. 1,270,375, and Taiwan Patent No. 2,7,556, the disclosure of which is incorporated herein by reference. The method mainly utilizes a simple and self-splicing method to protect the cell membrane of platelets, so that the specific sugar molecules present on the surface of the cell membrane are shielded, and the activity of the platelets can be maintained after being preserved for a period of time in vitro; the platelet-rich method of the method The blood supply may be the platelet concentration test from the blood center or the whole blood taken from the human body. (4) The method of separation. The blood is separated, and the plate is thick (4); this method can protect the platelets and maintain their activity in the bank. It can be stored in vitro for a long period of time, and there is no chemical residue to deposit the sputum material, so the contaminated platelets are not contaminated, the product is not picked up, and the activity remains intact, which can be used as a subject of in vitro study of its cell characteristics. Another method for in vitro preservation of platelets, red blood cells, non-nucleated cells and A-containing plate composition disclosed in Taiwan Patent Publication No. 200700556 is to provide a method for long-term preservation of blood plate. The steps include (a) anticoagulation a solution, a condensed precipitate, and a thrombin added to the physiological saline solution to form a solution; (6) adding the medium containing the platelet to the solution of the step (a) and forming a solution containing the platelet; and (6) 201004659 lyophilization step (b) The formed platelet solution is formed. Although the above two patent technologies respectively disclose a method for preserving liquid platelets and a method for preserving dry powdered platelets, the platelets used are taken from a mixture of blood donors such as blood centers, and are not directed to a specific individual. Platelets are preserved and therefore do not meet the needs of individual autologous platelet storage. SUMMARY OF THE INVENTION The present invention has been made in order to preserve an individual's own platelets for future use. SUMMARY OF THE INVENTION A primary object of the present invention is to provide a method for the dry preservation of autologous platelets to allow individuals to conveniently preserve autologous platelets. Another object of the present invention is to provide a method for the dry preservation of autologous platelets to enable individuals to easily advance themselves to the body and to improve their body. Still another object of the present invention is to provide a method for the self-contained platelet dry powdered storage method to allow individuals to withdraw money to prevent the use of fine platelets. For other purposes and functions of the present invention, please refer to the related form and the actual details, and the details are as follows. [Embodiment] Referring to the figure ^, the method for dry powder preservation of autologous platelets of the present invention is mainly for enabling a general individual (living body) to preserve autologous platelets, including the following steps: (a) Separating blood donation The machine draws single-frequency blood; 201004659 Take (four) ι物概鼓岭 (4) Touch the blood to separate the platelets for additional storage, and return the red blood cells and plasma to the living body; (C) Blood blood test for the blood to be examined, hospital routine Blood test (Routine bl (xxl tests) and other necessary money checks, and the storage of the arrival, recording, listening to the small board, bribe blood to protect her, maintain its activity; (4) judgment of the blood of the inspection (four) money If the county check is qualified, if the blood virus test fails, the platelet preservation operation is terminated (4); if the blood virus test is combined, the process proceeds to step (f), (1) the liquid-protected platelets are dry powdered to form an autologous dry powder. A small plate for the long-term preservation. The above-mentioned (a) (10) type of blood donation is difficult to obtain in the preparation of the living body for platelet intensive fluid or blood plate rich in gold plate. (4) The autologous dry powdered platelets are prepared by using platelets rich in blood virus-tested platelet concentrate or platelet-rich plasma rafts after the use of the living body for the surgery. The liquid-protected platelets in the above step (b) can be produced by the technique disclosed in the aforementioned Taiwan Patent No. 1,270,375, for example, including the following steps: • (1) First adding 5-30 ml of anticoagulant to 100 ml of physiological saline (N) 〇rmal sanne) 'Form a solution containing 5-30% anticoagulant, let stand for 5 minutes after shaking; (2) Add 〇. 5 5 5ml Cryoprecipitate; Cryo, fibrinogen in the above solution, After shaking, let stand for one minute; 8 201004659 (3) Add 0. 〇 (H-5ml of thrombin (Thrombin), shake and let stand for one minute; (4) Take platelets or try to contain platelet-containing plasma, Put in the above solution, let stand for ten minutes; (5) Store at 4 ° C or -20 ° C. • Or include the following steps: (1) First add 20ml anticoagulant to 100ml of physiological saline to form a solution containing 2% anticoagulant, shaken and then allowed to stand still 5 minutes; 〇 (2) add 〇·〇〇lml frozen precipitant in the above solution, shake and let stand for one minute; (3) add 5ml of thrombin, shake and let stand for one minute; (4) take platelet thick Liquid or platelet-rich plasma, placed in the above solution, allowed to stand for 10 minutes; (5) Store at -20 ° C. Or include the following steps: (1) First add 10ml anticoagulant to l〇〇mi In physiological saline, a solution containing 1% anticoagulant was formed, and it was allowed to stand for five minutes after shaking; (2) 3 ml of a frozen precipitant was added to the above solution, and the mixture was shaken and allowed to stand for one minute; (3) 0. 01 The thrombin was allowed to stand for one minute after shaking; (4) Take platelet thick liquid or platelet-rich plasma, put it into the above solution, and let it stand for ten minutes; (5) It can be stored under 4 it. Or include the following steps: 201004659 (1) First add 5ml anticoagulant to l〇〇mi physiological saline to form a solution containing 5% anticoagulant, shake and let stand for five minutes; (2) add 3ml of frozen precipitation agent In the above solution, shake and let stand for one minute; (3) add 0.0001 of thrombin, shake and let stand for one minute; (4) take platelet thick liquid or blood plate rich in gold plate, put into the above solution, Allow to stand for 10 minutes; (5) Store at 4 °C.抗 The anticoagulant used in the above step (1) may be an Acid Citrate Dextrose (ACD-A) or a Citrate Phosphate Dextrose (CPD). The autologous dry powdered platelets in the above step (f) can be produced by the technique disclosed in the aforementioned Taiwan Patent Publication No. 200700556, for example, including the following steps: (1) First adding 20 ml of ACD-A to 100 ml of physiological saline solution, Form a solution containing 2% anticoagulant, let stand for 5 minutes after shaking; ® (2) Add 〇. 〇〇lml condense the precipitate in the anticoagulant solution, shake and let stand for five minutes; (3) Add 5ml thrombin, let stand for 5 minutes after shaking; (4) Add platelet thick or platelet-rich plasma to the solution of step (3) and let stand for five minutes; (5) Finally freeze the solution of step (4) Store in powder form. The method for dry powder preservation of autologous platelets of the invention can achieve the following several effects: 201004659 1. Convenient preservation of the self-contained gray plate ^Invented can be a field of bribery from Wei powder dumping _ 2·Reducing waste and pollution of PRP Save for future improvement from (10) listening, Wei Cun _ up to two years to listen to ^ er delete the site preparation system, and - times, if the time is not used, it will be discarded, resulting in waste and pollution of PRP. By using the invention, the unused PRP can be stored and the powder growth factor can be preserved without any waste or pollution of pRp. 3. Reduce the frequency of user ash. Every time you need PRP, you need to collect blood. The user can save the autologous platelets at any time by using this month. It is not necessary to collect blood every time, which can reduce the blood collection rate. 4. Easy operation Everyone needs PRP, and the process of mining and centrifugation is required. It takes about 3 λ clock. The dry powder small plate made by the invention can be used after rehydration, and the operation is quite simple. 5. Saving costs. The PRP can be collected from the whole gold in the traditional way, and can only be used once. The cost per time needs 7500 yuan. 'If it is treated in the way of high dosage of the invention, it can be used 100 times each time. In the above, for a user who needs a medical cosmetic treatment for 2 or 3 months, 'the production of autologous dry powdered platelets can be used one or more times for postoperative maintenance, which saves money. The benefits are quite clear 201004659. The above description is only for the embodiments of the present invention, and any modifications and variations of the present invention will be apparent to those skilled in the art, and are not limited to the embodiments disclosed. 201004659 BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a flow chart showing a method for carrying out dry powdered preservation of autologous platelets of the present invention. [Description of main component symbols] (a), (b), (c), (d), (e), (f) are the number of steps for implementing the present invention.
1313