饲养层细胞

本文主要从饲养层细胞、无饲养层培养体系、培养基质、细胞因子等方面综述了hES细胞建系和维持其未分化状态的优化培养所取得的最新进展和存在的问题。

In this review , We will discuss the new progresses and unsolved issues on optimizing culture condi-tions for hES derivation and maintaining its undifferentiated state , mainly about feeder cell layer , feeder-free culture system , matrix and cytokines .

人角膜缘上皮细胞在丝裂霉素C处理的3T3饲养层细胞上形成边界清楚的克隆。

Results : Limbal epithelial cells showed clonal growth when cultured on mitomycin C treated3T3 fibroblasts feeder layer .

STO转基因小鼠饲养层细胞的最佳预处理条件

The best pretreatment condition of STO feeder layer cells in transgenic mice

一种利用STO饲养层细胞制备拟胚体的新方法

A Novel Method to form Embryoid Bodies Using STO Cells as Feeder Layers

CSCs饲养层细胞一般维持时间不超过8d；

The feeder layer cells of CSCs survived time did not exceed 8 d.

结果显示与支持细胞共培养能促进SSCs的存活和增殖，通过饲养层细胞辅助培养是体外培养SSCs较好的手段。

The results showed that with the support of co-cultured cells can promote the survival and proliferation of SSCs .

说明饲养层细胞在ES细胞的分化抑制中有一定作用，而起主导作用的是外源LIF。

These results indicate that feeder layer has certain action in suppressing ES cell differentiation in culture , while exogenous LIF play predominant role in maintaining ES cells undifferentiation . 4 .

选取生长旺盛的ESCs行碱性磷酸酶染色，克隆呈蓝紫色团块，而饲养层细胞不着色。

Selecting better growth of ESCs to do alkaline phosphatase staining , cell clones were blue-violet , and the feeder layer cells are not colored . 2 .

MEF细胞在第3代生长状态最好，最适合制备饲养层细胞。

MEF cells ' growth state in the 3rd generation is the best , which is the most suitable for preparing cells of feeder layer . 4 .

在成功分离、培养和鉴定人EG细胞的同时，比较了STO细胞和人胚胎肺成纤维细胞作为饲养层细胞对人EG细胞生长的影响。

Comparing influence on growth of human EG cells by STO and human embryonic lung fibroblasts respectively as feeder layers . ( 6 ) Finally , using soluble cytokines as the inducer to make human EG cells differentiate committed to hematopoietic cells .

结论：孕8、9周龄人胚为分离PGC的适宜材料，酶机械法分离PGC简单有效，饲养层细胞与生长因子为人PGC细胞体外培养所必需。

CONCLUSION : 8-and 9-week gestational age human embryo are optimal material for isolating human PGCs . Enzyme-mechanical method is simple and available to isolate human PGCs . Feeder layer and growth factors are necessary for human PGCs culture in vitro .

结果：（1）体外培养的小鼠ESC-D3细胞在胚胎成纤维饲养层细胞上连续传代培养，仍保持向3胚层分化的能力。

RESULTS : ( 1 ) The ESC-D3 cells kept the ability of differentiation into cellular derivations of all three primary germ layers after continuous passage culture .

昆明系小鼠饲养层细胞制备条件的优化

Optimization of Preparation Conditions for Feeder Layer Cells of Kunming Mice

以人皮肤成纤维细胞作为饲养层细胞培养人胚胎干细胞

Culture of human embryonic cells by using human dermal fibroblasts as feeder cells

转基因饲养层细胞体外促进脐带血CD34~+HSPC增殖

Transgenic cell line as a feeder layer on UCB CD34 ~ + HSPC in vitro amplification

不同饲养层细胞对胚胎干细胞和内细胞团的影响

Effect of Different Feeder Layers on Mouse Embryonic Stem Cells and Inner Cell Mass of Rabbit Blastocysts

饲养层细胞的接种密度对分离培养胚胎干细胞的影响

Effect of densities of feeder layer for isolation and culturing of Kunming white mouse embryonic stem cells

饲养层细胞体系及白血病抑制因子对维持小鼠胚胎干细胞未分化状态的作用

Effect of Feeder Layer Culture System and Leukemia Inhibitory Factor on Keeping Mouse Embryonic Stem Cells in Undifferentiation

然而，这种技术的应用通常需要依赖于饲养层细胞的支持，用于细胞培养的培养液通常也不能明确定位。

However , techniques for growing keratinocytes still often rely on feeder cell supports and culture medium that is not defined .

以自行分离、培养的人胚胎肺成纤维细胞为饲养层细胞，分离和培养了人胚胎生殖细胞，并进行鉴定。

Using human embryonic lung fibroblasts divided and cultivated by ourselves as the feeder layer to isolate and cultivate human embryonic germ cells and identify .

小鼠胎儿成纤维细胞经常作为饲养层细胞用于胚胎干细胞的培养。

Mouse embryonic fibroblast cells ( MEF ) are widely used as the feeder cells during the culture of embryonic stem cells ( ESCs ) .

目的寻求无血清、无饲养层细胞存在的情况下，胚胎干细胞向多巴胺能神经元定向诱导分化的最佳条件。

Objective To explore the optimal condition of direct differentiation into dopaminergic neurons of embryonic stem cells in serum-free and feeder layer cell free medium .

结论采用无饲养层细胞培养体系构建自体角膜上皮组织，移植后可以成功修复角膜缘干细胞缺失的角膜上皮。

Conclusion the reconstructed rabbit corneal epithelium hi vitro with a free-feeding layer culture system could be autografted to repair total corneal limbal stem cells deficiency .

目的采用两种方法制备饲养层细胞，对比观察培养效果，求得最佳培养方法，以期实现其有效利用。

Objective Adopt two methods to produce cells of feeder layer and compare and observe its result of culture to find a better method of culture .

通过两种方法来建立胎肝来源的细胞系：分离胎肝细胞挑取细胞克隆，与饲养层细胞共培养来建立细胞系；

In this study , we establish human fetal liver cell line by two ways : select the clonal fetal liver strains and coculture with feeder cells ;

为维持细胞在培养过程中的未分化状态，需要采用饲养层细胞培养，同时设计合理的培养液配方并添加多种抑制分化或促进增殖的细胞因子。

Feeder layer cells and suitable culture medium that supplemented with cytokines of promoting proliferation or restraining differentiation were crucial to keep chicken ES in an undifferentiated state .

同时，比较了饲养层细胞类型、生长因子、胚胎发育阶段等因素对绵羊胚胎干细胞分离培养效果的影响。

The effect of feeder cell type , growth factors , and embryo developmental stage on establishment of sheep embryonic stem cell was also studied in present experiment .

方法：分别采用人胚胎成纤维细胞和小鼠胚胎成纤维细胞为饲养层细胞，支持人受精卵的培养，观察其增殖和分化情况。

METHODS : Both mouse and human embryonic fibroblast cells were used as feeder layer to support human embryonic stem cells . The proliferation and differentiation of human embryonic cells were observed .

确定制备饲养层细胞的最佳方法。并利用荧光法检测制备的饲养层细胞有无支原体感染。

The best method of preparing feeder layer was determined by this method , and the cells of feeder layer were detected by fluorescence to test if it has been infected by Mycoplasma . 5 .

结论：在培养人EG细胞时，hELF饲养层较STO细胞饲养层更有优势。

( CONCLUSION : ) HELF as feeder layers is better than STO cells in culturing EG cells .