1283269 玖、發明說明 【發明所屬之技術領域】 本發明是有關於-種遺傳物質傳遞的方法，且特別是 有關於種細胞轉染（transfection)的方法。 【先前技術】 ^目前人類基因圖譜已經定序完成，有關基因方面的研 究也正式步入「後基因時代（p〇st_gen〇me era)」。在後基 因時代中，科學家的主要挑戰是在於要解開數萬基因代表 了那些蛋白質，而這些蛋白質之功能以及不同蛋白質之間 的關係為何。要解開這一連串問題的答案，最直接的方式 就是將想要瞭解的基因片段導入細胞中，讓細胞去表現這 段基因。 要將基因片段導入至真核細胞中，有各種不同之方 法’根據將外源核酸送入細胞内之載體來分，可將其分成 病毋型載體與非病毒型載體兩大類。使用病毒性載體之轉 染效率雖然高，但是所需技術層次高且製備程序繁複，不 易應用於大規模的轉染實驗中。使用非病毒性載體，其製 備方式簡易又迅速，較容易應用於大規模之轉染實驗中。 非病毒性載體例如有使用將近30年之磷酸鈣以及DEAe (diethyllaminoethyl)-葡聚糖（dextran)，還有較近發展出來 的帶正電荷之微脂粒（Liposome)等等。此三種化學試劑都 會和核酸形成複合物（complex)，以利核酸進入細胞中。 且上述二種方法之程序一般都是將含有核酸片段之轉染 1283269 以 試劑加入至具有細胞之培養皿或96井微量分析盤中， 進4亍核酸轉染的步驟。 最近 Ziauddin 與 Sabatini (Nature，2001，411: 107 與 USP 6,544,790)發展出一種逆轉染（reverse transfection) 方法。此逆轉染方法是先將外源質體核酸與明膠的混合物 直接點在上覆有P〇ly_L-lySine或是gamma-amin〇 propyl-silane之細胞培養平台上，然後加入以脂肪為主之 載體以覆蓋上述外源質體核酸與明膠的混合物之上一段 時間，以形成質體核酸-載體複合物。再將細胞加入至細 胞培養平台上，來進行核酸轉染的步驟。或是一開始即將 外源質體核酸、脂肪為主之載體與明膠混合，點在上覆有 P〇ly_L-iysine 或是 gamma韻in〇 pr〇pyi_sUane 之細胞培 養平台上。再將細胞加人至細胞培養平台上，來進行核酸 轉染的步驟。此種逆轉染技術可以應用來生產細胞微陣 列，以大量檢測不同基因片段之功能。 等之！若能大幅提昇轉染效率，應用在如細胞微陣列 ^刀析上，將可以同時對許多基因的 析。對於篩選特定蛋白質祚盔、Λ 進订刀 i m 作為〉口療樂物，或是要揭露去去 基因的功能，將會有十分大的助益。 ㈣路未知 【發明内容】 之 可 因此本發明的目的是在提供一種具有 細胞轉染方法。 木放率 ^月的另目的是在提供一種細胞轉染的方法， 1283269 應用於製造細胞微陣列。 裙據本&明之上述目的，提出一種細胞轉染的方法， 其步驟至少如下。先在細胞生長平台上形成一層能促進細 胞生長之水膠薄層。然後將轉染試劑點製在水膠薄層上之 多個轉染位置’此轉染試劑包含質體核酸與支鏈型聚 亞胺。接著’將細胞塗佈在水膠薄層上，讓位在轉染位置 上之細胞能接受轉染試劑中之質體核酸。 •根據本發明一較佳實施例，上述之水膠薄層之材質包 含天然水膠，例如瓊膠或明膠。 根據本發明另-較佳實施例，上述之轉染試劑還包含 穩定劑’如明膠、聚乙二醇或葡聚糖。 如上所述，應用本發明不僅可以簡易迅速地進行大量 之細胞轉染試驗’而^還可以得到高轉染效率之實驗結 果’對於最好使用微量材料來進行A量轉染實驗的細胞微 陣列來說’本發明提供了—個快速且可靠之細胞轉染方 法’以供探測可能之蛋白質新藥、可能之遺傳疾病的基因 以及基因在細胞内所扮演之功能。 根據上述’本發明提供一種細胞轉染（transfection) 的方法。因為非病毒性載體所傳送的基因在細胞中的表現 效率會父到細胞生長平台表面所塗佈材質的影響。而且外 源質體核酸的濃度以及在與載體形成複合物之後的複合 物穩定性’亦皆會影響到外源質體核酸之轉染效率。因 1283269 此’在本發明一較佳眚 貫知例中，探討了細胞生長平台表面 塗佈材質、轉染試劑中穩* 、 w τ穩又劑的種類與濃度以及外源核酸 /辰度對於細胞轉染效率的影塑。 在此較佳實施例φ，、@ m ^ ^ Α 1夕j中，選用含有綠色螢光蛋白基因之質 體核酸pEGFP-C 1作或从十好 1乍為外來基因，讓其轉染至中國大頰鼠 卵巢細胞（Chinese hamster ovary cell ； CHO cell ； ATCC number· CCL-61)中。因此，可以經由觀察ch〇細胞是否 有產生綠色螢光來確定質體核酸pEGFp_ci是否有轉染 至CHO細胞中。 實驗裎庠 先將含有綠色螢光蛋白基因之質體（plasmid)核酸 p_EGFP_Cl轉形進入宿主細胞大腸桿菌（dH_5〇;)中， 經過液態LB培養基（每升培養液中含有1〇克 bacto-tryptone、5 克 bacto-yeast 萃取物、10 克 NaCl)大 里培養之後’打破大腸桿菌細胞並使用Qiagen套組將質 體核酸pEGFP-Cl純化出來，再以超純水溶解核酸配成實 驗所需之質體核酸濃度。 在本較佳實施例中，選擇了載玻片為細胞生長平台， 細胞生長平台還可以為培養孤、微量分析盤或其他適合 者。接著在清洗載玻片之後，讓載玻片浸置在水膠 (hydrogel)溶液中約數分鐘。然後將載玻片取出，讓其乾 燥，讓水膠在載玻片上形成一層薄膜。然後將乾燥之水膠 塗佈玻片浸泡在70%乙醇中約5-15分鐘之久，以進行消 1283269 毒，再將消毒後之水膠塗佈玻片放入無菌操作台中乾燥後 備用。 上述所用之水膠溶液的濃度較佳為大於能讓其在載 玻片上形成一層薄膜的濃度。可用之水膠溶液包含天然水 膠溶液，例如重量百分比濃度大於1 %之瓊膠（agar)溶 液、大於1 %之壤爿曰凝膝（agarose)溶液或大於〇 2%之明 膠（gelatin B)溶液。 然後開始製備轉染試劑。將濃度約為〇·2 — 1.2 mg/mL 之支鏈型聚乙烯亞胺（polyethyleneimine ; pEI)溶液快速加 入至所需濃度之質體核酸溶液中，接著快速搖晃再靜置 20分鐘’使支鏈型聚乙烯亞胺與質體核酸能形成核酸複 合物，完成轉染試劑之製備。在製備轉染試劑時，亦可同 時加入不同之穩定劑。 再來’將玻片固定於手動式基因晶片製備台上，然後 以手動式基因晶片製備套組之針頭沾點上述之轉染試 劑’將轉染試劑點製在玻片上。之後，將點製完成的玻片 置於培養皿中。 加入適當數目的CHO細胞至動物細胞培養液中，使 其均勻地懸浮於培養液中，再直接將含CHO細胞之培養 液加在早先置於培養皿中的水膠塗佈玻片上方。經過24一 72小時培養之後，將玻片置於螢光顯微鏡下觀察，記錄 細胞轉染效率。 材質掛於細胞韓染效率之影释 1283269 在上述實驗程序中，塗佈載玻片所用之水膠溶液有重 曰 、 里百7刀比濃度分別為1 %之瓊膠溶液、1 %之瓊脂凝膠溶 液以及0.2%之明膠溶液。轉染試劑中之質體核酸濃度為 〇·45 # g/ # L ’且沒有加任何穩定劑。所得之實驗結果請 見表一 〇 表一：載玻片上所塗佈之水膠對於 效率之影響1283269 BRIEF DESCRIPTION OF THE INVENTION [Technical Field] The present invention relates to a method for the transmission of genetic material, and more particularly to a method for seed cell transfection. [Prior Art] ^ At present, the human gene map has been sequenced, and research on genes has officially entered the "post-gene era (p〇st_gen〇me era)". In the post-genogen era, the main challenge for scientists was to solve the tens of thousands of genes that represent those proteins, the function of these proteins and the relationship between different proteins. The most straightforward way to solve the answers to this series of questions is to introduce the gene fragments you want to know into the cells and let the cells express the genes. In order to introduce a gene fragment into a eukaryotic cell, there are various methods for dividing the exogenous nucleic acid into a cell, and dividing it into a diseased vector and a non-viral vector. Although the transfection efficiency using the viral vector is high, the required technical level is high and the preparation procedure is complicated, and it is not easy to be applied to large-scale transfection experiments. The use of non-viral vectors is simple and rapid, and is easier to apply to large-scale transfection experiments. Non-viral vectors include, for example, calcium phosphate and DEAe (diethyllaminoethyl)-dextran (dextran) which have been used for nearly 30 years, and more recently developed positively charged liposome (Liposome) and the like. All three chemical reagents form a complex with the nucleic acid to facilitate entry of the nucleic acid into the cell. Moreover, the procedures of the above two methods generally involve the step of transfecting the nucleic acid fragment 1283269 with a reagent into a petri dish with cells or a 96 well microanalytical tray for transfection of 4 亍 nucleic acid. Recently, Ziauddin and Sabatini (Nature, 2001, 411: 107 and USP 6, 544, 790) developed a reverse transfection method. The reverse transfection method is to directly place a mixture of exogenous plastid nucleic acid and gelatin directly on a cell culture platform covered with P〇ly_L-lySine or gamma-amin〇propyl-silane, and then add a fat-based carrier. To cover a mixture of the above exogenous plastid nucleic acid and gelatin for a period of time to form a plastid nucleic acid-carrier complex. The cells are then added to a cell culture platform for nucleic acid transfection. Or at the beginning, the exogenous plastid nucleic acid and fat-based carrier are mixed with gelatin and placed on a cell culture platform covered with P〇ly_L-iysine or gamma rhyme in pr〇pyi_sUane. The cells are then added to a cell culture platform for nucleic acid transfection. This reverse transfection technique can be applied to produce cell microarrays to detect the function of different gene segments in large quantities. Wait! If the transfection efficiency can be greatly improved, it can be applied to many genes at the same time, such as cell microarray analysis. It will be of great help to screen specific proteins, helmets, and knives as a singularity or to reveal the function of the gene. (4) Unknown road [Invention] It is therefore an object of the present invention to provide a method of transfection with a cell. Wood release rate Another purpose of the month is to provide a method of cell transfection, 1283269 applied to the manufacture of cell microarrays. Skirts According to the above objects of the present invention, a method of cell transfection is proposed, the steps of which are at least as follows. A thin layer of water gel that promotes cell growth is first formed on the cell growth platform. The transfection reagent is then spotted into a plurality of transfection sites on a thin layer of water gel. The transfection reagent comprises a plastid nucleic acid and a branched polyimine. The cells are then plated onto a thin layer of water gel so that the cells in the transfection site can receive the plastid nucleic acid in the transfection reagent. • According to a preferred embodiment of the invention, the material of the water-repellent layer comprises a natural water gel such as agar or gelatin. According to another preferred embodiment of the invention, the above transfection reagent further comprises a stabilizer such as gelatin, polyethylene glycol or dextran. As described above, the application of the present invention can not only perform a large number of cell transfection experiments in a simple and rapid manner, but also can obtain an experimental result of high transfection efficiency. For a cell microarray in which A-quantity transfection experiments are preferably performed using a trace amount of material. The present invention provides a fast and reliable method of cell transfection for detecting possible new protein drugs, genes for possible genetic diseases, and functions of genes within cells. According to the above, the present invention provides a method of cell transfection. Because the efficiency of the gene delivered by the non-viral vector in the cell is affected by the material applied to the surface of the cell growth platform. Moreover, the concentration of the exogenous plastid nucleic acid and the stability of the complex after complex formation with the carrier also affect the transfection efficiency of the exogenous plastid nucleic acid. In the preferred embodiment of the present invention, the type and concentration of the cell growth platform surface coating material, the transfection reagent, the w τ stabilizer, and the exogenous nucleic acid/length are discussed. The effect of cell transfection efficiency. In the preferred embodiment φ, @ m ^ ^ Α 1 夕 j, the plastid nucleic acid pEGFP-C 1 containing the green fluorescent protein gene is used as a foreign gene, and the gene is transfected to Chinese hamster ovary cells (CHO cell; ATCC number·CCL-61). Therefore, whether or not the plastid nucleic acid pEGFp_ci can be transfected into CHO cells can be determined by observing whether or not the ch〇 cells produce green fluorescence. In the experiment, the plasmid nucleic acid p_EGFP_Cl containing the green fluorescent protein gene was first transformed into the host cell Escherichia coli (dH_5〇;), and passed through a liquid LB medium (containing 1 gram of bacot-tryptone per liter of the culture solution). , 5 grams of bacto-yeast extract, 10 grams of NaCl) after culturing in E. coli cells and using the Qiagen kit to purify the plastid nucleic acid pEGFP-Cl, and then dissolving the nucleic acid in ultrapure water to prepare the assay. Body nucleic acid concentration. In the preferred embodiment, the slide is selected as a cell growth platform, and the cell growth platform can also be a cultured orphan, microanalytical tray or other suitable person. The slides were then immersed in a hydrogel solution for a few minutes after washing the slides. The slide is then removed and allowed to dry, allowing the water gel to form a film on the slide. Then, the dried water-coated slide glass is immersed in 70% ethanol for about 5-15 minutes to eliminate the poison of 1283269, and the disinfected water-coated slide glass is placed in an aseptic processing table and dried for use. The concentration of the aqueous gum solution used above is preferably greater than the concentration at which a film can be formed on the slide. The water gel solution that can be used comprises a natural water gel solution, such as an agar solution having a concentration of more than 1% by weight, a greater than 1% agarose solution or a gelatin B of greater than 2%. Solution. The preparation of the transfection reagent is then started. A solution of branched polyethyleneimine (pEI) at a concentration of about 〇·2 — 1.2 mg/mL is quickly added to the desired concentration of the plastid nucleic acid solution, followed by rapid shaking and rest for 20 minutes. The chain type polyethyleneimine and the plastid nucleic acid can form a nucleic acid complex, and the preparation of the transfection reagent is completed. When preparing the transfection reagent, different stabilizers may be added at the same time. Then, the slide was fixed on a manual type gene wafer preparation stage, and then the transfection reagent was spotted on the slide with a needle of a manual gene chip preparation kit dipped in the above-mentioned transfection reagent. After that, place the finished slide on the Petri dish. An appropriate number of CHO cells were added to the animal cell culture medium, and the suspension was uniformly suspended in the culture solution, and the CHO cell-containing culture solution was directly added over the hydrocolloid coated slide previously placed in the culture dish. After 24 to 72 hours of incubation, slides were placed under a fluorescent microscope and cell transfection efficiency was recorded. The material is hung on the cell dyeing efficiency. 1283269 In the above experimental procedure, the water gel solution used for coating the glass slide has a weight of 琼, 里, 7 knives, a concentration of 1% agar solution, 1% agar Gel solution and 0.2% gelatin solution. The concentration of the plastid nucleic acid in the transfection reagent was 〇·45 # g/ # L ' and no stabilizer was added. The experimental results obtained are shown in Table 1. 〇 Table 1: Effect of water gel coated on glass slides on efficiency
轉染效率（％)Transfection efficiency(%)
由表一可知，以載玻片上覆瓊脂凝膠所提供的細胞生 長壤境最差。與載破片上完全未塗佈水膠、塗料膠及塗 載玻片上塗佈之水膠 (重量百分) 無 相：’載玻片上覆璦脂凝膠所培育出的細胞數目只 有别述三者之1/3幻/4。對於轉染效率而 塗佈明膠之細胞轉染效率較佳，載玻片塗佈緩膠者載欠之， 載玻片塗佈璦賴膠者與載玻片完全未 染數目非常稀疏。 有之、、、田胞轉 穩疋劑對於細胞韓犖效率之影響 1283269 在上述實驗程序中，塗佈載玻片所用之水膠溶液有重 量百分比濃度分別為1%之瓊膠溶液、1%之瓊脂凝膠溶 液以及0.2°/。之明膠溶液。轉染試劑中之質體核酸濃度為 0·45 # g/ // L ’所加入之穩定劑有葡聚糖（dextran)、聚乙 二醇（Polyethylene Glycol ; PEG)以及明膠（gelatin B)，其 中上述之葡聚糖與聚乙二醇之分子量可約為1〇〇〇 _ 20000 〇 在此所用之葡聚糖之分子量分別約為1 5 〇〇以及 15000 — 20000，重量百分比濃度分別為〇%、〇.25%、〇 5% 與1%，其實驗結果請分別參考第丨圖與第2圖。所用之 聚乙二醇之分子量約為2000，重量百分比濃度分別為 ㈣、0.25%、0.5%與1%’其實驗結果請參考第3圖。而 所用之明膠的重量百分幻農度分別為0%、〇.〇125%、 0.025%與0.05%，其實驗結果請參考第4圖。 在第卜4圖中，可以發現在轉染試劑中加入穩定劑葡 聚糖與聚乙二醇之後，上面塗佈有1%之瓊膠溶液、1%之 瓊脂凝膠溶液以及0.2%之明膠唆饬★莽χ L古 5膠，合液之载玻片的細胞轉 :效率均較沒有加入穩定劑之前要來得高。若以第i圖斑 ΓΛ之：驗結果來比較，亦可發現分子量約為·之 葡聚糖對於細胞轉染效率之提升比八 、 15000-20000之葡聚糖的效果要好。刀子量約為 ^, 由上逑可知，鱗定劑 對於、、、田胞轉染效率之提升有彳艮大的幫助。 心 !283269 在上述實驗程序中，塗佈載玻片所用之 量百分屮：塗麻达 乂 /分饮馬重 /又為〇·2%之明膠溶液。轉染試劑中之質體校 酸巧分別為。.”、、…與…—：： 之穩定劑有分子旦 里約為15〇〇〇 — 20000之葡聚糖以及八 薏約為2000之奄7 社 次刀子 為0% ' —_ 4聚糖之重量百分比濃度分別 、曲 .5/°、〇·5%與1。/。，而聚乙二醇之重量百分比 漢度^別為0%、0.25%、05%與1%。 百刀比 實紅果睛見第5_6 gj，其中第5圖以分子 15000 - 20000之不同濃度的葡聚糖作為穩定劑，在不同 質體核酸濃度下進行細胞轉染所得之細胞轉染效率圖，而 第6圖以分子量約為2〇〇〇之不同濃度的聚乙二醇作為穩 疋劑’在不同質體核酸濃度下進行細胞轉染所得之細胞轉 染效率圖。 在第5圖中，其所用之穩定劑為分子量約為15〇〇〇_ 2000之葡聚糖對於細胞轉染效率有提升之效果。當其重 量百分比濃度4 〇·25%時，有較佳之細胞轉染效率，所 得之細胞㈣效率約為5Q%左右。而且細胞轉染效率大 致隨著質體核酸濃度之增加而增加，在質體核酸之濃度為 0.45 // g/# L時有最佳之細胞轉染效率。 在第6圖中，其所用之穩定劑為分子量約為2〇⑻之 聚乙二醇對於細胞轉染效率有提升之效果。當其重量百分 比7辰度為0 · 5 %時，有較Y击由：九 ^权佳之細胞轉染效率，所得之細胞 轉染效率約為70%左右。而且細胞轉染效率大致隨著質 體核酸漠度之增加而增加，在f體核酸之濃度為〇 3〜 10 1283269 # L時有最佳之細胞轉染效率。 由上述本發明較佳實施例可知，在這些較佳實施例中 Μ $微陣列打點器在塗佈水膠之細胞生長平台上形成微 米大j之孔/同，並直接注入奈升（nan〇 liter)之轉染試劑。 然後在此水膠表面上培養動物細胞，讓轉染試劑中之核酸 可以進入動物細胞中。經過一段時間之後，動物細胞就可 以表現出轉染試劑中核酸的基因。 因此應用本發明，不僅可以簡易迅速地進行大量之細 胞轉染試驗，而且還可以得到高轉染效率（高達7G%)之實 驗結果。對於最好使用微量材料來進行大量轉染實驗的細 胞微陣列來說，本發明提供了 一個快速且可靠之細胞轉毕 方法，以供探測可能之蛋白質新藥、可能之遺傳疾病的基 因以及基因在細胞内所扮演之功能。 雖然本發明已以一較佳實施例揭露如上，然1並非用 以限定本發明，任何熟習此技藝者，在不脫離本發明之精 =範圍内’當可作各種之更動與潤飾，因此本發明之保 濩範圍當視後附之申請專利範圍所界定者為準。 【圖式簡單說明】 本發明所附圖式之詳細說明如下： 第^圖是以分子量約為1500之不同濃度的葡聚糖作 ^穩定劑，在有不同水膠塗佈之載破片上進行細胞轉染所 件之細胞轉染效率圖。 1283269 第2圖是以分子量約為15〇〇〇 _ 2〇〇〇〇之不同濃度 葡聚糖作為較劑，在有不同水膠塗佈之載二田 胞轉染所得之細胞轉染效率圖。 進仃、、、田 :二圖是以分子量約為2_之不同濃 作錢定劑，在有不同水膠塗佈之載玻片上進行細胞轉; 所知之細胞轉染效率圖。 、 ^ 4圖是以不同濃度的明膠作為穩定劑，在有不同水 膠塗佈之載玻片上進行細胞轉染所As can be seen from Table 1, the cell growth gradient provided by the slide-coated agar gel was the worst. The water gel (weight percent) coated on the fully uncoated water gel, the coating glue and the coated glass slide on the fragment is phaseless: 'The number of cells grown on the slide coated with the rouge gel is only three 1/3 illusion / 4. For the transfection efficiency, the transfection efficiency of the gelatin-coated cells is better, and the slide-coated slow-adhesive is insufficient, and the number of the glass-coated smear-coated gels and the slides is completely unstained. The effect of turf, turf, and sputum on the efficiency of cell 荦 128 1283269 In the above experimental procedure, the water gel solution used for coating slides has a concentration of 1% agar solution, 1% by weight. Agar gel solution and 0.2 ° /. Gelatin solution. The concentration of plastid nucleic acid in the transfection reagent is 0.45 #g/ // L 'The stabilizers added are dextran, polyethylene glycol (PEG) and gelatin B. The molecular weight of the above-mentioned glucan and polyethylene glycol may be about 1 〇〇〇 20000 〇 The molecular weight of the dextran used here is about 15 〇〇 and 15000 2020, respectively, and the concentration by weight is 〇. %, 〇.25%, 〇5% and 1%, please refer to the figure and figure 2 for the experimental results. The molecular weight of the polyethylene glycol used was about 2,000, and the concentrations by weight were (4), 0.25%, 0.5%, and 1%, respectively. For the experimental results, please refer to Figure 3. The weight percent of the gelatin used is 0%, 〇.〇125%, 0.025% and 0.05%, respectively. For the experimental results, please refer to Figure 4. In Fig. 4, it can be found that after adding the stabilizer dextran and polyethylene glycol to the transfection reagent, the above is coated with 1% agar solution, 1% agar gel solution and 0.2% gelatin.唆饬★莽χ L Gu 5 gel, cell transfer of the slides of the mixture: the efficiency is higher than before the stabilizer is added. If the results are compared with the results of the i-th image, it can be found that the glucan with a molecular weight of about ‧ is better than the dextran of eight and 15,000-20000 for the improvement of cell transfection efficiency. The amount of knives is about ^. As can be seen from the top, the sizing agent has greatly helped the improvement of the transfection efficiency of the field. Heart !283269 In the above experimental procedure, the amount of sputum used to apply the slide: 麻 达 乂 / 饮 重 重 又 又 又 2 2 2 2 2 2 2 2 2 2 。 。. The plastids in the transfection reagents are as follows. ",,... and...-:: Stabilizers have a molecular weight of about 15 〇〇〇 - 20,000 glucan and gossip about 2000 奄 7 knives are 0% ' - _ 4 glycans The weight percentages are respectively ±5°°, 〇·5% and 1%, and the weight percentage of polyethylene glycol is 0%, 0.25%, 05% and 1%. See the 5th 6th gj of the real red fruit, and the 5th figure shows the transfection efficiency of the cells obtained by cell transfection at different plastid nucleic acid concentrations with different concentrations of dextran from 15000 to 20000 as the stabilizer. The graph shows the transfection efficiency of cells obtained by cell transfection at different plastid nucleic acid concentrations using polyethylene glycol with a molecular weight of about 2 不同 as a stabilizer. In Figure 5, it is used. The stabilizer is a glucan with a molecular weight of about 15 〇〇〇 _ 2000, which has an effect of improving the transfection efficiency of the cells. When the concentration is 4 〇·25% by weight, the cell transfection efficiency is better, and the obtained cells (4) The efficiency is about 5Q%, and the cell transfection efficiency increases roughly with the increase of plastid nucleic acid concentration. The optimal cell transfection efficiency is 0.45 // g/# L. In Figure 6, the stabilizer used is a polyethylene glycol with a molecular weight of about 2 〇 (8) for cell transfection efficiency. There is an improvement effect. When the weight percentage of 7 is 0. 5 %, there is a Y transfection efficiency: the transfection efficiency of the cell is about 70%, and the cell transfection efficiency is about 70%. The efficiency generally increases with the increase of the plastid nucleic acid inversion, and the optimal cell transfection efficiency is obtained when the concentration of the f-form nucleic acid is 〇3 to 10 1283269 # L. From the above preferred embodiments of the present invention, these are In the preferred embodiment, the microarray indicator forms a micron-sized hole/same on the cell growth platform coated with water gel, and directly injects a nano-liter transfection reagent. The animal cells are cultured on the surface, and the nucleic acid in the transfection reagent can enter the animal cells. After a period of time, the animal cells can express the genes of the nucleic acid in the transfection reagent. Therefore, the present invention can be used not only to perform a large amount of simple and rapid Cell transfection test And can also obtain experimental results with high transfection efficiency (up to 7G%). For cell microarrays that preferably use trace materials for bulk transfection experiments, the present invention provides a fast and reliable method for cell transfer For the detection of possible protein new drugs, genes of possible genetic diseases, and functions of genes in cells. Although the invention has been disclosed above in a preferred embodiment, 1 is not intended to limit the invention, A person skilled in the art can make various changes and modifications without departing from the scope of the invention, and the scope of the invention is defined by the scope of the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS The detailed description of the drawings of the present invention is as follows: The first figure is a dextran having a molecular weight of about 1500 as a stabilizer, which is carried out on a fragment of a hydrogel coated with different water gel. Transfection efficiency map of cells transfected. 1283269 Fig. 2 is a graph showing the transfection efficiency of cells transfected with different water gels coated with different concentrations of dextran at a molecular weight of about 15 〇〇〇 2 〇〇〇〇. . Into the 仃, , , , : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : , ^ 4 Figure is a different concentration of gelatin as a stabilizer, cell transfection on different hydrogel coated glass slides
^ ς 曰 于之、、、田胞轉染效率圖。 第5圖以/刀子$約為15〇〇〇、 聚糖作為敎劑，在不同質體核酸濃产下^同濃度的葡 得之細胞轉染效率圖。 行細胞轉染所 不同濃度的聚乙二醇作 進行細皰轉染所得之細 第6圖以分子量約為2〇〇〇之 為穩定劑，在不同質體核酸濃度下 胞轉染效率圖。^ ς 于 Yu Zhi,,, Tian cell transfection efficiency map. Figure 5 shows the transfection efficiency of the same concentration of glucosinolates in the same concentration of different plastid nucleic acids with a knife of about 15 〇〇〇 and a polysaccharide as an elixir. Cellular transfection of different concentrations of polyethylene glycol for fine blister transfection. Figure 6 shows a cell transfection efficiency at different plastid nucleic acid concentrations with a molecular weight of approximately 2 稳定 as a stabilizer.
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