北京生物醫(yī)學(xué)工程

胚胎干細(xì)胞與聚3-羥基丁酸酯-co-4-羥基丁酸酯共培養(yǎng)細(xì)胞補(bǔ)片的初步研究

Primary study on 3-hydroxybutyrate-co-4-hydroxybutyrate co-cultured with mouse embryonic stem cell of cell patch

作者：馬義祥穆軍升張健群伯平

單位：首都醫(yī)科大學(xué)附屬北京安貞醫(yī)院 (北京100029)

關(guān)鍵詞：胚胎干細(xì)胞；細(xì)胞補(bǔ)片；聚3-羥基丁酸酯-co-4-羥基丁酸酯；三維培養(yǎng)

分類號(hào)：R318.04

出版年·卷·期（頁(yè)碼）：2016·35·1（53-58）

摘要：

目的干細(xì)胞移植是具有治療心血管疾病潛力的有效方法，但是細(xì)胞移植過(guò)程中缺乏有效的載體會(huì)大大影響其成活和增殖。聚3-羥基丁酸酯-co-4-羥基丁酸酯P(3HB-co-4HB)是一種能夠完全降解的，具有良好物理特性的高分子材料。探討P(3HB-co-4HB)這種三維材料是否適合胚胎干細(xì)胞在上面生長(zhǎng)，并生成細(xì)胞補(bǔ)片。方法復(fù)蘇小鼠胚胎干細(xì)胞，傳代培養(yǎng)，消化離心后重懸細(xì)胞，用懸滴法形成擬胚體(embryid bodies,EB)，將擬胚體與P(3HB-co-4HB)膜共培養(yǎng)；72h共培養(yǎng)后固定并進(jìn)行掃描電鏡和共聚焦顯微鏡觀察細(xì)胞黏附情況，DAPI熒光染色觀察細(xì)胞在P(3HB-co-4HB)膜上形態(tài)。結(jié)果胚胎干細(xì)胞在培養(yǎng)皿中貼壁生長(zhǎng)良好，形態(tài)正常；擬胚體同P(3HB-co-4HB)膜共培養(yǎng)72h后，所有擬胚體都在三維材料表面良好生長(zhǎng)，爬膜率為100%；Confocal顯微鏡和掃描電鏡觀察到在2mm的三維膜上，擬胚體生長(zhǎng)貼附在生物膜上，細(xì)胞形態(tài)正常。結(jié)論 ESC在P(3HB-co-4HB)表面生長(zhǎng)、增殖形成細(xì)胞補(bǔ)片。P(3HB-co-4HB)可作為干細(xì)胞治療多種疾病的支架材料之一。

Objective Stem cell transplantation is considered promising for treating cardiovascular disorders. However,lack of an effective carrier impacts their survival and proliferation. Poly 3-hydroxybutyrate-co-4-hydroxybutyrate ［P(3HB-co-4HB)］ is a novel polymer biomaterial with excellent physical characteristics. We assessed the suitability of P(3HB-co-4HB) combined with embryonic stem cells (ESC) for cell patch formation. Methods Mouse embryonic stem cells were recovered，passaged，digested，centrifuged，suspended and form embryoid bodies (EB)by hanging drop method.EB were cultured with P(3HB-co-4HB) together for 72 hours. The morphology of EBs attached on the films was observed by confocal and scanning electron microscopy. DAPI fluorescence staining to observe cell morphology. Results Adherent mouse ESC displayed normal growth and morphology.When cultured as EB for 72 hours,100% grew on the biofilm and cells were present in the 3D biofilm to a depth of 2mm,with normal cell morphology. Conclusions Polymer biomaterial patches composed of P(3HB-co-4HB) permit ESCs growth .P(3HB-co-4HB) is a suitable biomaterial for stem cell transplantation in the treatment of cardiovascular disorders.

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