|
|
|
|
|
|
| The role of aqueous humor in the proliferation of retinal progenitor cells |
|
| 作者：杨静, Mo… 文章来源：北京大学第三医院眼科中心 点击数1593 更新时间：2005/6/28 21:03:08 文章录入：yjbell 责任编辑：毛进 |
|
|
|
| Purpose:
To examine pig aqueous humor for stimulatory activity on embryonic rat retinal progenitor cells (RPCs) in vitro and partly to identify the components that contribute to this activity.
Methods:
RPCs was prepared from the retina of embryonic day 20 Sprague Dawley rats and were cultured in medium consisting of DMEM:F12 together with N2-supplement, epidermal growth factor(EGF), basic fibroblast growth factor (bFGF), leukemia inhibitory factor (LIF), in the presence or absence of aqueous humor for 0 to 4 days. Cell proliferation and cluster formation was quantified by 3H-thymidine assay and morphometric analysis, respectively. Antibodies used to characterize the RPCs included markers for neural progenitors (Nestin and Sox-2) and astrocytes (glial fibrillary acidic protein [GFAP] ). by the use of flow cytometry. Active fractions of porcine aqueous humor was identified by the use of size exclusion chromatograghy (ÄktaExploreSystem)
Results:
Addition of aqueous humor to the culture medium enhanced within 4 days both the number and size of clusters formed with 66% and 29%, respectively (p<0.01). This was further evident by an increased cell proliferation demonstrated with 3H-thymidine assay (p<0.01). By use of gelfiltration chromatography it was possible to demonstrate both high molecular fractions 30 – 40 kDa and low molecular fractions < 1 kDa containing the growth promoting activities. 22%, and 16% of the primary cells was found to express Nestin and Sox-2, respectively. GFAP was not detected..
Conclusions:
The results demonstrate that aqueous humor contain factors which can stimulate formation and growth of RPC clusters in vitro. By size-exclusion gelchromtography it is possible to identify fractions with specific growth promoting activity.
|
|
|
|
|
|
|
|
|
|
