骨髓基质细胞和CD34+的造血祖细胞间的交流由connexin43型通道连接调节

hxf_86

Intercellular communication between bone marrow stromal cells and CD34+ haematopoietic progenitor cells is mediated by connexin 43-type gap junctions
: k7 i' h* R: m# Q7 G* I; o骨髓基质细胞和CD34+的造血祖细胞间的交流由connexin43型通道连接调节
( s, T7 g: E5 U# k+ a! [' s1 L& W
6 l4 ^3 I- {5 Q" P摘要：( N" l4 b/ K; C
关于造血祖细胞（HPC）和基质细胞间是否存在功能性的通道连接，一直存在着争议。原始的CD34+的HPC与人类骨髓基质细胞系L87/4共培养。利用具有高度敏感性的全细胞钳技术，可以发现大多数的（91%）CD34+的HPC与L87/4间有电流交流。更重要的是，）CD34+的HPC依附在L87/4后的1小时内可以观察到有效的偶联。通过对比，由L87/4组成的同类细胞间有显著更高的电偶联。单通道导电率分析发现了一种形式，以connexin43型通道连接为特征，对于同类和不同类的细胞都是这样的。这个Cx表型通过Cx-43特异性的单克隆抗体流式细胞仪分析和RT-PCR的验证。最后，染料转染实验利用降落伞技术使得染料扩散而不影响浆膜成分，这个实验是对电生理实验的补充。总的来说，我们的数据表明，功能性的Cx-43型通道连接存在于基质细胞和未成熟的造血祖细胞中，可能提供造血过程中一个重要的调节通道。6 Q; }8 f+ ]& n( a5 h: ?

$ A& A0 v; r! o; i# X2 u9 YSummary. The existence of functional gap junctions between haematopoietic progenitor cells (HPCs) and stromal
$ P% j( Z- o7 x/ f$ F2 |cells of the haematopoietic microenvironment in the human system is a controversial issue. Primary CD341 HPCs5 D2 E# G, N- a  @0 R+ W
isolated from leukapheresis products were co-incubated with the human fibroblastoid bone marrow stromal cell* n! c' B8 F" T4 N3 Q  g! G
line L87/4 in short-term liquid culture. Using the highly sensitive double whole-cell patch-clamp technique, we
; \( A) J. `9 }3 Cfound that the majority (91%) of CD341 HPCs are electrically coupled to L87/4 cells. Importantly, efficient! L4 e: {7 `1 u, t) j
coupling was observed within 1 h of the attachment ofCD341 HPCs to plastic adherent L87/4 cells. By comparison,
' A; |1 F, L$ A: b8 shomologous cell pairs formed by L87/4 cells exhibited a significantly higher electric coupling. Analysis of singlechannel conductances revealed an electric profile characteristic of connexin 43 (Cx43)-type gap junctions for both homologous and heterologous cell pairs. The Cx phenotype was confirmed using Cx43-specific monoclonal antibodies in a flow cytometric assay and reverse transcription polymerase chain reaction (RT-PCR) for the detection of Cx43 mRNA. Finally, the electrophysiological studies were complemented by dye-transfer experiments using the recently described `parachute' technique that allows the monitoring of dye diffusion without disruption of the plasma membrane. Taken together, our data indicate that functional Cx43-type gap junctions exist between stromal cells and immature HPCs and, thus, may provide an important regulatory pathway in haematopoiesis.$ c9 v3 w% ?5 J1 \% ]& C9 @

suning5

may be

细胞海洋

请按照翻译小组的版规修改一下你的文章