CN110894466A - 一种干细胞培养用培养基 - Google Patents

一种干细胞培养用培养基 Download PDF

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CN110894466A
CN110894466A CN201911322538.3A CN201911322538A CN110894466A CN 110894466 A CN110894466 A CN 110894466A CN 201911322538 A CN201911322538 A CN 201911322538A CN 110894466 A CN110894466 A CN 110894466A
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梁勇
张首平
谢海博
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Guangzhou Hairun Kanghua Biotechnology Co Ltd
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Abstract

本发明公开了一种干细胞培养用培养基，包括底部护盖，所述底部护盖的内腔设有培养基本体，所述底部护盖的顶部啮合有顶部护盖，所述底部护盖顶部的两侧以及顶部护盖底部的两侧开设有相互配合使用的扣槽，所述底部护盖和顶部护盖的使用材质均为亚克力材料，所述养基本体包括有15%FBS，无水氯化钙116.6mg/L，L‑亮氨酸59.05mg/L，亚油酸0.042mg/L，无水硫酸铜0.0013mg/L，L‑赖氨酸盐酸盐91.25mg/L，硫辛酸0.105mg/L，九水硝酸铁0.05Lmg/L，L‑蛋氨酸17.24mg/L，酚红8.1mg/L，七水硫酸亚铁0.417mg/L，L‑苯丙氨酸35.48mg/L，1，4‑丁二胺二盐酸盐0.081mg/L。本发明制备方法简单，使用材料仪器少，可大量制配后用以培养使用，干细胞的扩增速度快，干细胞分化能力强，可分化成多种功能细胞，具有很高的科研和医学应用价值。

Description

一种干细胞培养用培养基
技术领域
本发明涉及干细胞培养技术领域，具体为一种干细胞培养用培养基。
背景技术
干细胞是21世纪生物医学界普遍关注的新研究课题，干细胞是一类具有自我复制能力的多潜能细胞，在一定条件下，它可以分化成多种功能细胞，根据干细胞所处的发育阶段分为胚胎干细胞和成体干细胞，根据干细胞的发育潜能分为三类：全能干细胞、多能干细胞和单能干细胞，干细胞是一种未充分分化，尚不成熟的细胞，具有再生各种组织器官和人体的潜在功能，医学界称为“万用细胞”，干细胞培养基是作用于干细胞的培养过程中所需要的特殊培养环境，以达到干细胞的贴壁或增殖效果。
为提升干细胞培养的效率，我们提出一种干细胞培养用培养基的制配方法。
发明内容
本发明的目的在于提供一种干细胞培养用培养基，具备制备简单，便于工作人员配制，繁殖效果好的优点，解决了现有市场上干细胞培养制配方法较为繁琐，培养过程费时费力的问题。
为实现上述目的，本发明提供如下技术方案：一种干细胞培养用培养基，包括底部护盖，所述底部护盖的内腔设有培养基本体，所述底部护盖的顶部啮合有顶部护盖。
优选的，所述底部护盖顶部的两侧以及顶部护盖底部的两侧开设有相互配合使用的扣槽，所述底部护盖和顶部护盖的使用材质均为亚克力材料。
优选的，所述养基本体包括有15%FBS，无水氯化钙116.6mg/L，L-亮氨酸59.05mg/L，亚油酸0.042mg/L，无水硫酸铜0.0013mg/L，L-赖氨酸盐酸盐91.25mg/L，硫辛酸0.105mg/L，九水硝酸铁0.05Lmg/L，L-蛋氨酸17.24mg/L，酚红8.1mg/L，七水硫酸亚铁0.417mg/L，L-苯丙氨酸35.48mg/L，1，4-丁二胺二盐酸盐0.081mg/L，氯化钠6999.5mg/L，L-天门冬酰胺7.5mg/L，氯化胆碱8.98mg/L，L-精氨酸盐酸盐147.5mg/L，L-脯氨酸17.25mg/L，盐酸吡哆醛2mg/L，L-异亮氨酸54.47mg/L，次黄嘌呤2mg/L，维生素B120.68mg/L，白血病抑制因子5～10μg/L；碱性成纤维细胞生长因子5～10μg/L和蒸馏水。
优选的，所述培养基本体的制备方法为：
A、将锥形瓶、试管、培养皿、量筒等没入含有洗涤剂的水中，用毛刷洗，然后用自来水及蒸馏水冲净，移液管先用含有洗涤剂的水浸泡，再用自来水及蒸馏水冲洗，洗刷干净的玻璃器皿置于烘箱中烘干后备用。
B、将称好的药品置于一烧杯中，先加人少量蒸馏水，用玻璃棒搅动，加热溶解。
C、将配制好的溶液密封放入灭菌锅进行高温杀菌，在0.105MPa压力下，温度121℃时，灭菌15～30min即可。
D、将溶液放入培养容器中，加入待培养的干细胞密封放入37°C，含有百分之五的CO2的加湿培养箱中培养。
与现有技术相比，本发明的有益效果如下：
通过底部护盖和顶部护盖的透明材料使用，能够更加方便使用者在培养过程中及时对培养情况进行查看，通过底部护盖和顶部护盖表面扣槽的开设，能够使两者之间密封性更好，有效避免在密封的过程中外界的细菌进入影响培养效果，不利于使用者的培育使用。
本发明制备方法简单，使用材料仪器较少，可大量制配后用以培养使用，干细胞的扩增速度快，干细胞分化能力强，可分化成多种功能细胞，具有很高的科研和医学应用价值。
附图说明
图1为本发明结构示意图。
图中：1、底部护盖；2、培养基本体；3、顶部护盖。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
在发明的描述中，需要说明的是，术语“上”、“下”、“内”、“外”“前端”、“后端”、“两端”、“一端”、“另一端”等指示的方位或位置关系为基于附图所示的方位或位置关系，仅是为了便于描述本发明和简化描述，而不是指示或暗示所指的装置或元件必须具有特定的方位、以特定的方位构造和操作，因此不能理解为对本发明的限制。此外，术语“第一”、“第二”仅用于描述目的，而不能理解为指示或暗示相对重要性。
在发明的描述中，需要说明的是，除非另有明确的规定和限定，术语“安装”、“设置有”、“连接”等，应做广义理解，例如“连接”，可以是固定连接，也可以是可拆卸连接，或一体地连接；可以是机械连接，也可以是电连接；可以是直接相连，也可以通过中间媒介间接相连，可以是两个元件内部的连通。对于本领域的普通技术人员而言，可以具体情况理解上述术语在本发明中的具体含义。
本发明的底部护盖1、培养基本体2和顶部护盖3部件均为通用标准件或本领域技术人员知晓的部件，其结构和原理都为本技术人员均可通过技术手册得知或通过常规实验方法获知。
请参阅图1，一种干细胞培养用培养基，包括底部护盖1，底部护盖1的内腔设有培养基本体2，底部护盖1的顶部啮合有顶部护盖3。
底部护盖1顶部的两侧以及顶部护盖3底部的两侧开设有相互配合使用的扣槽，底部护盖1和顶部护盖3的使用材质均为亚克力材料。
培养基本体2包括有15%FBS，无水氯化钙116.6mg/L，L-亮氨酸59.05mg/L，亚油酸0.042mg/L，无水硫酸铜0.0013mg/L，L-赖氨酸盐酸盐91.25mg/L，硫辛酸0.105mg/L，九水硝酸铁0.05Lmg/L，L-蛋氨酸17.24mg/L，酚红8.1mg/L，七水硫酸亚铁0.417mg/L，L-苯丙氨酸35.48mg/L，1，4-丁二胺二盐酸盐0.081mg/L，氯化钠6999.5mg/L，L-天门冬酰胺7.5mg/L，氯化胆碱8.98mg/L，L-精氨酸盐酸盐147.5mg/L，L-脯氨酸17.25mg/L，盐酸吡哆醛2mg/L，L-异亮氨酸54.47mg/L，次黄嘌呤2mg/L，维生素B120.68mg/L，白血病抑制因子5～10μg/L；碱性成纤维细胞生长因子5～10μg/L和蒸馏水。
培养基本体2的制备方法为：
A、将锥形瓶、试管、培养皿、量筒等没入含有洗涤剂的水中，用毛刷洗，然后用自来水及蒸馏水冲净，移液管先用含有洗涤剂的水浸泡，再用自来水及蒸馏水冲洗，洗刷干净的玻璃器皿置于烘箱中烘干后备用。
B、将称好的药品置于一烧杯中，先加人少量蒸馏水，用玻璃棒搅动，加热溶解。
C、将配制好的溶液密封放入灭菌锅进行高温杀菌，在0.105MPa压力下，温度121℃时，灭菌15～30min即可。
D、将溶液放入培养容器中，加入待培养的干细胞密封放入37°C，含有百分之五的CO2的加湿培养箱中培养。
使用时，本发明制备方法简单，使用材料仪器较少，可大量制配后用以培养使用，干细胞的扩增速度快，干细胞分化能力强，可分化成多种功能细胞，具有很高的科研和医学应用价值。
尽管已经示出和描述了本发明的实施例，对于本领域的普通技术人员而言，可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型，本发明的范围由所附权利要求及其等同物限定。

Claims (4)

1.一种干细胞培养用培养基，包括底部护盖（1），其特征在于：所述底部护盖（1）的内腔设有培养基本体（2），所述底部护盖（1）的顶部啮合有顶部护盖（3）。
2.根据权利要求1所述的一种干细胞培养用培养基，其特征在于：所述底部护盖（1）顶部的两侧以及顶部护盖（3）底部的两侧开设有相互配合使用的扣槽，所述底部护盖（1）和顶部护盖（3）的使用材质均为亚克力材料。
3.根据权利要求1所述的一种干细胞培养用培养基，其特征在于：所述培养基本体（2）包括有15%FBS，无水氯化钙116.6mg/L，L-亮氨酸59.05mg/L，亚油酸0.042mg/L，无水硫酸铜0.0013mg/L，L-赖氨酸盐酸盐91.25mg/L，硫辛酸0.105mg/L，九水硝酸铁0.05Lmg/L，L-蛋氨酸17.24mg/L，酚红8.1mg/L，七水硫酸亚铁0.417mg/L，L-苯丙氨酸35.48mg/L，1，4-丁二胺二盐酸盐0.081mg/L，氯化钠6999.5mg/L，L-天门冬酰胺7.5mg/L，氯化胆碱8.98mg/L，L-精氨酸盐酸盐147.5mg/L，L-脯氨酸17.25mg/L，盐酸吡哆醛2mg/L，L-异亮氨酸54.47mg/L，次黄嘌呤2mg/L，维生素B120.68mg/L，白血病抑制因子5～10μg/L；碱性成纤维细胞生长因子5～10μg/L和蒸馏水。
4.根据权利要求1所述的一种干细胞培养用培养基，其特征在于：所述培养基本体（2）的制备方法为：
A、将锥形瓶、试管、培养皿、量筒等没入含有洗涤剂的水中，用毛刷洗，然后用自来水及蒸馏水冲净，移液管先用含有洗涤剂的水浸泡，再用自来水及蒸馏水冲洗，洗刷干净的玻璃器皿置于烘箱中烘干后备用；
B、将称好的药品置于一烧杯中，先加人少量蒸馏水，用玻璃棒搅动，加热溶解；
C、将配制好的溶液密封放入灭菌锅进行高温杀菌，在0.105MPa压力下，温度121℃时，灭菌15～30min即可；
D、将溶液放入培养容器中，加入待培养的干细胞密封放入37°C，含有百分之五的CO2的加湿培养箱中培养。
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